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1 DOI: /ncb2697 Figure S1 Cytokeratin 5 is a specific marker for basal and intermediate cells in all mouse prostate lobes. (a) Immunofluorescence staining showing co-localization of YFP with p63 in basal cells (arrowheads) of tamoxifeninduced CK5-CreER T2 ; R26R-YFP/+ anterior prostate (AP). (b) Lack of co-localization of YFP with the luminal marker CK18. (c,d) Co-localization of YFP with CK5 in the dorsolateral prostate (DLP, c) and ventral prostate (VP, d). (e-i) Overlay (e) and individual channels (f-i) demonstrating that all p63-positive cells (arrowheads) are also CK5-positive, and 99.1% of CK5- positive cells are also p63-positive (n=3,132/3,159, 3 animals analyzed). (j-n) Overlay (j) and individual channels (k-n) demonstrating that all CK14- positive cells (arrowheads) are CK5-positive, and vice versa. (o-s) Overlay (o) and individual channels (p-s) showing lineage-marking (YFP-positivity) of an intermediate cell (arrow) that is p63-positive and CK18-positive. Scale bars correspond to 50 microns. 1
2 Figure S2 Marker analyses of basal cells isolated by flow cytometry and of the resulting spheres and renal grafts. (a-l) Purity of basal cells isolated by flow cytometry as shown in Fig. 1b, followed by immunofluorescence staining of cytospin preparations. (a-d) Overlay (a) and individual channels (b-d) showing co-localization of YFP and CK5 in the isolated basal cell population. (e-h) Overlay (e) and individual channels (f-h) showing colocalization of YFP and p63. (i-l) Overlay (i) and individual channels (j-l) showing absence of CK18 expression in the isolated basal cell population. (m-r) Spheres generated from basal cells after 7 days treatment with DHT, showing expression of luminal (CK18) and basal (p63, CK5) markers. (s) Quantitation of sphere formation from YFP + Lin Sca-1 + CD49f hi cells (8.0% box in Fig. 1c), compared to the remaining YFP + cells. 5 replicates from the same sorted population were performed for each group, and two sample t-tests were used for statistical analysis; error bars correspond to standard deviation and show variabilities beween replicates. (t) Detection of synaptophysin-positive neuroendocrine cell (arrow) in renal graft from basal cells. Scale bars in a-r correspond to 50 microns and in t to 100 microns; error bars in s correspond to one standard deviation. 2
3 Figure S3 Analysis of cell proliferation during prostate regeneration. (a) Time course of prostate regression and regeneration in tamoxifentreated CK5-CreER T2 ; R26R-YFP/+ mice. (b-e) Ki67 immunostaining to detect cell proliferation in CK5-positive basal cells (arrowheads) (b,d) and CK18-positive luminal cells (c,e) at 2 days after androgen administration to initiate regeneration (b,c) or 4 days after androgen administration (d,e). (f) CK5-CreER T2 ; R26R-YFP/+ mice were treated with BrdU during the first 12 days of prostate regeneration. (g,h) Immunostaining for BrdU incorporation to detect cell proliferation in CK5-positive basal cells (arrowhead, g) and CK18-positive luminal cells (h) at full regeneration. Scale bars in b-e,g-h correspond to 50 microns. 3
4 Figure S4 Additional data for lineage-tracing analysis of basal epithelial cells. (a) Alternative protocol for lineage-tracing during prostate regeneration, in which tamoxifen induction for marking of basal cells occurs in the regressed state. (b) Specific co-localization of YFP with CK5-expressing basal cells (arrowheads) after tamoxifen induction in androgen-deprived CK5-CreER T2 ; R26R-YFP/+ anterior prostate. (c-h) Most YFP + cells express the basal marker CK5 (arrowheads, c,e,g) and not the luminal marker CK18 (panels d,f,h) after regeneration in the anterior prostate (AP, c,d), ventral prostate (VP, e,f), and dorsolateral prostate (DLP, g,h), although rare YFP + CK18 + luminal cells (arrows, d,f,h) can be detected in all three lobes. (i-m) Lineage-tracing of basal cells during regeneration in older mice. (i) Strategy for lineage-tracing during prostate regeneration in mice starting at 10 months of age. (j,k) Co-localization of YFP with CK5-expressing basal cells (arrowheads) after regeneration in CK5-CreER T2 ; R26R-YFP/+ DLP (j) or AP (k). (l) General absence of YFP and CK18 co-localization in anterior prostate. (m) Detection of a rare YFPexpressing CK18-positive luminal cell (arrow) in anterior prostate. Scale bars correspond to 50 microns. 4
5 Figure S5 Phenotypes of PIN/tumor lesions following Pten deletion in basal cells. (a) Strategy for analysis of PIN/tumor lesion formation at six months after tamoxifen induction of hormonally-intact CK5-CreER T2 ; Pten flox/flox ; R26R-YFP/+ mice. (b-g) Whole-mounts of dissected anterior prostate (AP, b,e), dorsolateral prostate (DLP, c,f), and ventral prostate (VP, d,g) lobes from control uninduced CK5-CreER T2 ; Pten flox/flox ; R26R- YFP/+ mice (b-d) or tamoxifen induced mice of the same genotype (e-g). (h,i) Hematoxylin-eosin staining of PIN/tumor lesions in the DLP (h) and VP (i). (j,k) Widespread phosphorylated Akt (pakt) expression at 6 months after induction in the DLP (j) and VP (k). Scale bars in b-g correspond to 2 mm, in h,i to 100 microns, and in j,k to 50 microns. 5
6 Figure S6 Phenotypes of PIN/tumor lesions following Pten deletion in luminal cells. (a) Time course for tumor formation after tamoxifen induction of hormonally-intact Nkx3.1 CreERT2/+ ; Pten flox/flox ; R26R-YFP/+ mice. (b-d) Hematoxylin-eosin staining of anterior prostate (AP, b,c,e) and dorsolateral prostate (DLP, d) showing low-grade PIN at 1 month after induction (b), high-grade PIN at 3 months after induction (c,d), and high-grade PIN/ carcinoma in situ at 6 months after induction (e). (f,g) Widespread phosphorylated Akt (pakt) expression after three months of induction of Nkx3.1 CreERT2/+ ; Pten flox/flox ; R26R-YFP/+ mice (f) is comparable to the pakt immunostaining observed after six months of induction of CK5-CreER T2 ; Pten flox/flox ; R26R-YFP/+ mice (basal origin, g) (h-n) Marker analysis at three months after induction of Nkx3.1 CreERT2/+ ; Pten flox/flox ; R26R-YFP/+ mice shows Ki67 immunoreactivity (arrow, h), lack of CK5 expression (i), expression of CK18 (j) and AR (k), and existence of CK5 + CK18 + intermediate cells (l-n). (o) Expression of selected genes of the luminal origin leading-edge signature in three different human patient datasets 22, 26, 28 in the Oncomine database, showing their up-regulation in human prostate cancer (dark blue boxplots) compared with benign prostate (light blue boxplots). Scale bars in b-e correspond to 100 microns, and in f-n to 50 microns. 6
7 Supplementary Tables Table S1 Supporting data for quantitation of basal cell properties. Table S2 Summary of histopathological phenotypes in mice with PIN/tumor lesions of basal or luminal origin. Table S3 List of 200 differentially expressed genes between Basal 3 (3 months after induction) and Basal control (uninduced). Table S4 List of 200 differentially expressed genes between Luminal 1 (1 month after induction) and Luminal control (uninduced). Table S5 List of 200 differentially expressed genes between Basal 6 (6 months after induction) and Basal 3 (3 months after induction). Table S6 List of 200 differentially expressed genes between Luminal 3 (3 months after induction) and Luminal 1 (1 month after induction). Table S7 List of 200 differentially expressed genes between Luminal 3 (3 months after induction) and Basal 6 (6 months after induction). Table S8 List of 200 differentially expressed genes between High-risk (death within 12 months) and Low-risk (survival for more than 192 months) human patient samples. Table S9 List of biological pathways enriched in the Luminal 3 vs Basal 6 signature. Table S10 List of genes in the leading edge from comparison of mouse Luminal 3 vs. Basal 6 signature with human high vs. low risk lethality signature. Table S11 Antibodies used in this study. 7
AP VP DLP H&E. p-akt DLP
A B AP VP DLP H&E AP AP VP DLP p-akt wild-type prostate PTEN-null prostate Supplementary Fig. 1. Targeted deletion of PTEN in prostate epithelium resulted in HG-PIN in all three lobes. (A) The anatomy
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Page 1 09/10/2013 AD Award Number: W81XWH-12-1-0453 TITLE: The cytoplasm translocation of the androgen receptor cofactor p44 as a target for prostate cancer treatment PRINCIPAL INVESTIGATOR: Zhengxin Wang
Supplementary Fig. 1 Blocking shh function at the protein level confirms its role as a guidance cue for postcommissural axons.
Supplementary Fig. 1 Blocking shh function at the protein level confirms its role as a guidance cue for postcommissural axons. As an alternative method to demonstrate the role of shh as a guidance cue
Supplementary Figure 1. A. Bar graph representing the expression levels of the 19 indicated genes in the microarrays analyses comparing human lung
Supplementary Figure 1. A. Bar graph representing the expression levels of the 19 indicated genes in the microarrays analyses comparing human lung immortalized broncho-epithelial cells (AALE cells) expressing
Supplemental Table S1
Supplemental Table S. Tumorigenicity and metastatic potential of 44SQ cell subpopulations a Tumorigenicity b Average tumor volume (mm ) c Lung metastasis d CD high /4 8. 8/ CD low /4 6./ a Mice were injected
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DOI: 10.1038/ncb2535 Figure S1 SOX10 is expressed in human giant congenital nevi and its expression in human melanoma samples suggests that SOX10 functions in a MITF-independent manner. a, b, Representative
M2 microglia/ macrophages drive oligodendrocyte differentiation during CNS remyelination
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Supplementary Figure 1. EC-specific Deletion of Snail1 Does Not Affect EC Apoptosis. (a,b) Cryo-sections of WT (a) and Snail1 LOF (b) embryos at
Supplementary Figure 1. EC-specific Deletion of Snail1 Does Not Affect EC Apoptosis. (a,b) Cryo-sections of WT (a) and Snail1 LOF (b) embryos at E10.5 were double-stained for TUNEL (red) and PECAM-1 (green).
SSM signature genes are highly expressed in residual scar tissues after preoperative radiotherapy of rectal cancer.
Supplementary Figure 1 SSM signature genes are highly expressed in residual scar tissues after preoperative radiotherapy of rectal cancer. Scatter plots comparing expression profiles of matched pretreatment
TITLE: Prostate Expression Databases: Gene Expression Resources for Comparative Studies of Prostate Carcinogenesis
AD Award Number: W81XWH-05-1-0110 TITLE: Prostate Expression Databases: Gene Expression Resources for Comparative Studies of Prostate Carcinogenesis PRINCIPAL INVESTIGATOR: Peter S. Nelson, Ph.D. CONTRACTING
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DOI: 10.1038/ncb3200 Supplementary Figure 1 Expression analysis of stomach markers in gutlike structure. (a) Differentiation scheme of gut-like structure formation from embryonic stem cells. (b) RT-PCR
PHENOTYPIC DYNAMICS OF MICROGLIAL AND MONOCYTE-DERIVED CELLS IN GLIOBLASTOMA-BEARING MICE.
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Supplementary Information
Supplementary Information Title Degeneration and impaired regeneration of gray matter oligodendrocytes in amyotrophic lateral sclerosis Authors Shin H. Kang, Ying Li, Masahiro Fukaya, Ileana Lorenzini,
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SUPPLEMENTARY INFORMATION Human cerebral cortex development from pluripotent stem cells to functional excitatory synapses Yichen Shi 1,2, Peter Kirwan 1,2, James Smith 1,2, Hugh P.C. Robinson 3 and Frederick
CHAPTER VII CONCLUDING REMARKS AND FUTURE DIRECTION. Androgen deprivation therapy is the most used treatment of de novo or recurrent
CHAPTER VII CONCLUDING REMARKS AND FUTURE DIRECTION Stathmin in Prostate Cancer Development and Progression Androgen deprivation therapy is the most used treatment of de novo or recurrent metastatic PCa.
Supplementary Figure 1
d f a IL7 b IL GATA RORγt h HDM IL IL7 PBS Ilra R7 PBS HDM Ilra R7 HDM Foxp Foxp Ilra R7 HDM HDM Ilra R7 HDM. 9..79. CD + FOXP + T reg cell CD + FOXP T conv cell PBS Ilra R7 PBS HDM Ilra R7 HDM CD + FOXP
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SUPPLEMENTARY INFORMATION Supplementary Figure 1. The expression of ephrin-b2 H2BGFP persists in the post-hearingonset organ of Corti and is specifically restricted to supporting cells. Sox2 immunolabeling
Supplemental Information. Myocardial Polyploidization Creates a Barrier. to Heart Regeneration in Zebrafish
Developmental Cell, Volume 44 Supplemental Information Myocardial Polyploidization Creates a Barrier to Heart Regeneration in Zebrafish Juan Manuel González-Rosa, Michka Sharpe, Dorothy Field, Mark H.
Supplementary Figure 1. Electroporation of a stable form of β-catenin causes masses protruding into the IV ventricle. HH12 chicken embryos were
Supplementary Figure 1. Electroporation of a stable form of β-catenin causes masses protruding into the IV ventricle. HH12 chicken embryos were electroporated with β- Catenin S33Y in PiggyBac expression
Supplementary Materials for VAMP4 directs synaptic vesicles to a pool that selectively maintains asynchronous neurotransmission
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Supplementary Figure 1
Supplementary Figure 1 Localization of virus injections. (a) Schematic showing the approximate center of AAV-DIO-ChR2-YFP injection sites in the NAc of Dyn-cre mice (n=8 mice, 16 injections; caudate/putamen,
Figure S1. (A) Schematic diagram of dnrar transgene allele. (B) X-Gal staining of testis from
Figure S1. (A) Schematic diagram of dnrar transgene allele. (B) X-Gal staining of testis from germ cell mutants (dnrar flox/flox, Stra8-Cre +, RARElacZ) (A ), controls (dnrar flox/flox, RARElacZ) (B ),
Supplemental Figure 1. Quantification of proliferation in thyroid of WT, Ctns -/- and grafted
Supplemental Figure 1. Quantification of proliferation in thyroid of WT, Ctns -/- and grafted Ctns -/- mice. Cells immunolabeled for the proliferation marker (Ki-67) were counted in sections (n=3 WT, n=4
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DOI: 10.1038/ncb3021 Supplementary figure 1 Characterisation of TIMPless fibroblasts. a) Relative gene expression of TIMPs1-4 by real time quantitative PCR (RT-qPCR) in WT or ΔTimp fibroblasts (mean ±