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1 Electronic Supplementary Material (ESI) for Integrative Biology. This journal is The Royal Society of Chemistry 2018 Supplementary Material 1
2 Supplemental Figures Supplemental Figure S1. Mechanical properties of collagen gels. Collagen gels were measured using a rheometer with a 40 mm aluminum cone (2 degrees) geometry. The angular frequency was 1 Hz and measurements were taken every 1 minute for a duration of 1 hour. The G and G values were taken as an average of 10 data points between 50 minute and 60 minutes. 2
3 Supplemental Figure S2. Images of untreated cells and corresponding mitochondria in 1-4 mg/ml collagen. a-b) Cell in 1 mg/ml collagen with images of brightfield and mcherry c-d) Cell in 2 mg/ml collagen with images of brightfield and mcherry e-f) Cell in 3 mg/ml collagen with images of brightfield and mcherry g-h) Cell in 4 mg/ml collagen with images of brightfield and mcherry. Scale bar is 10 µm. 3
4 Supplemental Figure S3. DMSO treatment of cells. a) MSD curves of MDA-MB-231 cells treated with DMSO and control cells embedded in 1 mg/ml collagen. b) β curves of of MDA-MB- 231 cells treated with DMSO and control cells embedded in 1 mg/ml collagen. c) MSD curves of MDA-MB-231 cells treated with DMSO and control cells embedded in 2 mg/ml collagen. d) β curves of of MDA-MB-231 cells treated with DMSO and control cells embedded in 2 mg/ml collagen e) MSD curves of MDA-MB-231 cells treated with DMSO and control cells embedded in 3 mg/ml collagen. f) β curves of of MDA-MB-231 cells treated with DMSO and control cells embedded in 3 mg/ml collagen g) MSD curves of MDA-MB-231 cells treated with DMSO and control cells embedded in 4 mg/ml collagen. h) β curves of of MDA-MB-231 cells treated with DMSO and control cells embedded in 4 mg/ml collagen 4
5 Supplemental Figure S4. Blebbistatin treated cells. Cells were incubated with blebbistatin (50 µm) for 2 hours in 1-4 mg/ml collagen gels. a) MSD curves of MDA-MB-231 cells treated with blebbistatin embedded in 1-4 mg/ml collagen. b) β curves of of MDA-MB-231 cells treated with blebbistatin embedded in 1-4 mg/ml collagen 5
6 Supplemental Figure S5. Comparison of Blebbistatin treated cells with control cells. a) MSD curves of MDA-MB-231 cells treated with blebbistatin and control cells embedded in 1 mg/ml collagen. b) β curves of of MDA-MB-231 cells treated with blebbistatin and control cells embedded in 1 mg/ml collagen. c) MSD curves of MDA-MB-231 cells treated with blebbistatin and control cells embedded in 2 mg/ml collagen. d) β curves of of MDA-MB-231 cells treated with blebbistatin and control cells embedded in 2 mg/ml collagen e) MSD curves of MDA-MB-231 cells treated with blebbistatin and control cells embedded in 3 mg/ml collagen. f) β curves of of MDA-MB-231 cells treated with blebbistatin and control cells embedded in 3 mg/ml collagen g) MSD curves of MDA- MB-231 cells treated with blebbistatin and control cells embedded in 4 mg/ml collagen. h) β curves of of MDA-MB-231 cells treated with blebbistatin and control cells embedded in 4 mg/ml collagen 6
7 Supplemental Figure S6. Comparison of untreated with drug treated cells. a) Comparison of MSDs at t = 50 ms of untreated and drug treated cells in 1-4 mg/ml collagen. b) Comparison of β s at t = 1 s of untreated and drug treated cells in 1-4 mg/ml collagen. Blebbistatin and DMSO treated conditions are also shown for comparison. Error bars are s.e.m. * indicates p<
8 Supplemental Figure S7. Relationship between collagen concentration and effective cell stiffness. a) Power output generated by molecular motors of cells embedded in 1-4 mg/ml collagen b) Shear modulus of cells embedded in 1-4 mg/ml collagen. Assumed mitochondria with radius r tracer = 70 nm. 8
9 Supplemental Figure S8. Cell morphology analysis of untreated, GM6001, Y27632, and Paclitaxel treated cells in 1-4mg/mL collagen. a) Cell area b) Cell perimeter c) Aspect ratio, and d) Circularity of phallodidin stained cells. Number of cells for untreated (1 mg/ml n=15, 2 mg/ml n=21, 3 mg/ml=15, 4 mg/ml n=15), GM6001 treated cells (1 mg/ml n=14, 2 mg/ml n=22, 3 mg/ml n=15, 4 mg/ml n=15), Y27632 treated cells (1 mg/ml n=15, 2 mg/ml n=22, 3 mg/ml n=15, 4 mg/ml n=14), and Paclitaxel treated cells (1 mg/ml n=15, 2 mg/ml n=23, 3 mg/ml n=15, 4 mg/ml n=15). Error bars are s.e.m. 9
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