Figure S1. Reduction in glomerular mir-146a levels correlate with progression to higher albuminuria in diabetic patients.
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1 Supplementary Materials Supplementary Figures Figure S1. Reduction in glomerular mir-146a levels correlate with progression to higher albuminuria in diabetic patients. Figure S2. Expression level of podocyte markers Nephrin and WT1 are reduced in the kidney sections of diabetic mice. Figure S3. Expression level of mir-146a and its targets in primary podocytes. Figure S4. WB analysis of ErbB4 expression level in primary podocytes and murine kidney sections. Figure S5. Schematic of the luciferase reporter assay. Figure S6. Ectopic expression of mir-146a results in reduced ErbB4 expression in murine podocytes. Figure S7. STZ treatment increases renal leukocyte infiltration that is suppressed by erlotinib. Figure S8. ErbB4 and EGFR heterodimerize in cells.
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3 Figure S1. Reduction in glomerular mir-146a levels correlate with progression to higher albuminuria in diabetic patients. A-B. Two graphs showing change in urinary albumin to creatinine ratio (ACR) in diabetic patients (n=45) over 5±1 years (T 1 = initial ACR measurement, at the time of kidney biopsy, T 2 = second ACR measurement 5±1 years later), where patients with relative glomerular mir-146a levels of >0.4 show slower increase in ACR over this time period and patients with relative mir-146a levels of <0.4 show progression to a higher level of ACR over the same period. Each dot represents an individual patient. Significance of difference between the two sets was determined using student s t-test.
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5 Figure S2. Expression level of podocyte markers Nephrin and WT1 are reduced in the kidney sections of diabetic mice. Bar graphs showing relative expression levels of Nephrin and WT1 in the kidney sections from 12 wk old BTBR WT and BTBR ob/ob mice, as measured by qrt-pcr. Their expression was normalized with Gapdh mrna expression. Data shown are mean ± SEM (n=3). *, P<0.05.
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7 Figure S3. Expression level of mir-146a and its targets in primary podocytes. A. A bar graph showing relative expression level of mir-146a in the kidney sections from wk old WT and mir- 146a -/- mice, as measured by qrt-pcr. Their expression was normalized using mir-361 mirna expression. Data shown are mean ± SEM (n=3). *, P<0.05. B. Bar graphs showing relative expression level of mir-146a targets ErbB4 and Notch-1 in the primary podocytes isolated from the glomeruli of wk old WT and mir-146a -/- mice, as measured by qrt-pcr. Their expression was normalized using GAPDH mrna expression. Expression level of EGFR, not a mir-146a target, was not changed. Data shown are mean ± SEM (n=3). **, P<0.01, *, P<0.05.
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9 Figure S4. WB analysis of ErbB4 expression level in primary podocytes and murine kidney sections. ErbB4 levels are basally increased in mir-146a -/- samples. Immunoblot analysis of total ErbB4 protein in the lysates from isolated glomeruli of WT or mir-146a -/- (-/-) animals (left blot) or lysates of cultured primary podocytes from isolated glomeruli of WT and mir-146a -/- (-/-) (right blot). β-actin was used as the loading control. Relative position of the molecular weight markers is shown on the left.
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11 Figure S5. Schematic of the luciferase reporter assay. A. Schematic graph of luciferase reporter assay. Luciferase expression plasmid contains 3 -UTR sequence of our interest at the 3 -end of luciferase gene. Transfection of the luciferase expression plasmid into HEK293T cells produces the luciferase mrna with 3 -UTR of our interest. In the absence of a specific mir-146a sequence to suppress the 3 -UTR, the luciferase protein product is produced, which can be detected using a luciferase assay. However, in the presence of a co-transfected mir-146a construct, and if this construct recognizes the inserted 3 -UTR sequence, the translation of luciferase mrna is suppressed and the expression and activity of luciferase is reduced, which can be measured using a luciferase assay system. B. Sequence of the wide type and mutated 3 -UTRs of ErbB4 and Notch1 that were inserted in the luciferase vector are listed.
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13 Figure S6. Ectopic expression of mir-146a results in reduced ErbB4 expression in murine podocytes. Immunoblot analysis of total ErbB4 protein in the lysates from cultured WT podocytes transduced with viral particles containing plasmids that express either mir-146a or a scrambled sequence. GAPDH was used as the loading control. Relative position of the molecular weight markers is shown on the left.
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15 Figure S7. STZ treatment increases renal leukocyte infiltration that is suppressed by erlotinib. Diabetic milieu increases inflammation and tissue infiltration of leukocytes. Representative immunohistochemical images of anti-cd11b antibody stained murine kidney sections from WT (left panels) and mir-146a -/- (right panels) mice treated with vehicle alone (Control), with STZ and vehicle (STZ) or with STZ and erlotinib (STZ + Erl) at the end of the experiment (16 weeks post-stz) and show increased leukocyte infiltration in the STZ-treated tissues, which is reduced in the STZ and erlotinib group. Images were taken at 40X magnification. Scale bar, 50 µm. A bar graph below shows quantification of CD11b positive cells in four independent, randomly selected fields observed at 10X magnification. Data shown are mean ± SEM (n=3-5/grp). *, P<0.05; **, P<0.01.
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17 Figure S8. ErbB4 and EGFR heterodimerize in cells. A. HEK293T cells were transfected with the plasmids expressing either EGFR or human ErbB4 and EGFR (as indicated) and cultured for 2 days. Subsequently, the lysates were immunoprecipitated with antibody against ErbB4 or an isotype control according to literature protocols. The immunoprecipitated extracts were immunoblotted (IB) for EGFR or ErbB4 using specific antibodies to study co-immunoprecipitation of ErbB4 with EGFR. Total cell lysates were used as a control to confirm expression of each protein. Relative position of the molecular weight markers is shown on the left.
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Supplementary Information Titles Journal: Nature Medicine Article Title: Corresponding Author: Supplementary Item & Number Supplementary Fig.1 Fig.2 Fig.3 Fig.4 Fig.5 Fig.6 Fig.7 Fig.8 Fig.9 Fig. Fig.11
Quantitative PPARγ expression affects the balance between tolerance and immunity
Quantitative PPARγ expression affects the balance between tolerance and immunity Ya-Hui Liu 1, Yau-Sheng Tsai 1,2,3, Shih-Chieh Lin 4, Nan-Shih Liao 5, Ming-Shiou Jan 6, Chung-Tiang Liang 7, Shih-Wen Hsu
Supplementary Figure 1. SC35M polymerase activity in the presence of Bat or SC35M NP encoded from the phw2000 rescue plasmid.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 Supplementary Figure 1. SC35M polymerase activity in the presence of Bat or SC35M NP encoded from the phw2000 rescue plasmid. HEK293T
Dynamic Interaction of Stress Granule, DDX3X and IKK-α Mediates Multiple Functions in
Dynamic Interaction of Stress Granule, and Mediates Multiple Functions in Hepatitis C Virus Infection Véronique Pène, Qisheng Li#, Catherine Sodroski, Ching-Sheng Hsu, T. Jake Liang# Liver Diseases Branch,
Supplementary Figures
Supplementary Figures Supplementary Figure 1 DOT1L regulates the expression of epithelial and mesenchymal markers. (a) The expression levels and cellular localizations of EMT markers were confirmed by
Supplementary Figure 1. PD-L1 is glycosylated in cancer cells. (a) Western blot analysis of PD-L1 in breast cancer cells. (b) Western blot analysis
Supplementary Figure 1. PD-L1 is glycosylated in cancer cells. (a) Western blot analysis of PD-L1 in breast cancer cells. (b) Western blot analysis of PD-L1 in ovarian cancer cells. (c) Western blot analysis
Supplementary Fig. 1. Delivery of mirnas via Red Fluorescent Protein.
prfp-vector RFP Exon1 Intron RFP Exon2 prfp-mir-124 mir-93/124 RFP Exon1 Intron RFP Exon2 Untransfected prfp-vector prfp-mir-93 prfp-mir-124 Supplementary Fig. 1. Delivery of mirnas via Red Fluorescent
SUPPLEMENTARY FIGURES
SUPPLEMENTARY FIGURES Figure S1. Clinical significance of ZNF322A overexpression in Caucasian lung cancer patients. (A) Representative immunohistochemistry images of ZNF322A protein expression in tissue
SUPPLEMENTARY INFORMATION
DOI: 1.138/ncb222 / b. WB anti- WB anti- ulin Mitotic index (%) 14 1 6 2 T (h) 32 48-1 1 2 3 4 6-1 4 16 22 28 3 33 e. 6 4 2 Time (min) 1-6- 11-1 > 1 % cells Figure S1 depletion leads to mitotic defects
T H E J O U R N A L O F C E L L B I O L O G Y
Supplemental material Díaz et al., http://www.jcb.org/cgi/content/full/jcb.201209151/dc1 T H E J O U R N A L O F C E L L B I O L O G Y Figure S1. Hypoxia induces invadopodia formation in different epithelial
Nature Immunology: doi: /ni Supplementary Figure 1
Supplementary Figure 1 NLRP12 is downregulated in biopsy samples from patients with active ulcerative colitis (UC). (a-g) NLRP12 expression in 7 UC mrna profiling studies deposited in NCBI GEO database.
DOI: 10.1038/ncb2210 b. ICAM1 ng ml -1 P = 0.0001 Small RNA (15-30nts) ng ml -1 Cell Lysate Exosome HDL Plasma HDL Normal Human HDL mirnas R = 0.45 P < 0.0001 Normal Human Exosome mirnas Figure S1. Characterization
L1 on PyMT tumor cells but Py117 cells are more responsive to IFN-γ. (A) Flow
A MHCI B PD-L1 Fold expression 8 6 4 2 Fold expression 3 2 1 No tx 1Gy 2Gy IFN Py117 Py117 Supplementary Figure 1. Radiation and IFN-γ enhance MHCI expression and PD- L1 on PyMT tumor cells but Py117 cells
Lentiviral Delivery of Combinatorial mirna Expression Constructs Provides Efficient Target Gene Repression.
Supplementary Figure 1 Lentiviral Delivery of Combinatorial mirna Expression Constructs Provides Efficient Target Gene Repression. a, Design for lentiviral combinatorial mirna expression and sensor constructs.
Intracellular MHC class II molecules promote TLR-triggered innate. immune responses by maintaining Btk activation
Intracellular MHC class II molecules promote TLR-triggered innate immune responses by maintaining Btk activation Xingguang Liu, Zhenzhen Zhan, Dong Li, Li Xu, Feng Ma, Peng Zhang, Hangping Yao and Xuetao