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D ay 0 1 2 3 4 5 6 7 Viable C ellN um ber 0 10 5 2x10 5 3x10 5 4x10 5 5x10 5 6x10 5 C ontrol 1,25D3 BAF CQ S4 Online Resource 3 S5/S6 Sensitivity to chloroquine and bafilomycin A1 ZR- 75-1 cells were exposed to various doses of Bafilomycin A1 and Chloroquine and toxicity was assessed by the MTT assay 72 hours following initial drug treatment. Adriamycin was used as a positive control (S5). Cells were exposed to 100nM 1,25D3, 200nM Bafilomycin A1 or 5µM Chloroquiune and viable cell number was assessed by trypan blue exclusion at days 3 and 6 post treatment (S6) S7 Quantification of AVOs . Percentage of cells with positive AVO staining was monitored using flow cytometry at 72 hours post treatment. Values shown are from a representative experiment with triplicate samples for each condition #p<0.05 compared to IR. ##p<0.05 compared to 1,25D3+IR nline Resource 3 A bsorbance (M TT) 0.0 0.1 0.2 0.3 0.4 0.5 S5 S6 P ercen t P o sitive A V O S taining 0 10 20 30 40 # ## S7
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Online Resource 3. S5. S4. S6. S7. Online Resource 3 - PowerPoint PPT Presentation
Transcript of Online Resource 3
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S5/S6 Sensitivity to chloroquine and bafilomycin A1 ZR-75-1 cells were exposed to various doses of Bafilomycin A1 and Chloroquine and toxicity was assessed by the MTT assay 72 hours following initial drug treatment. Adriamycin was used as a positive control (S5). Cells were exposed to 100nM 1,25D3, 200nM Bafilomycin A1 or 5µM Chloroquiune and viable cell number was assessed by trypan blue exclusion at days 3 and 6 post treatment (S6) S7 Quantification of AVOs. Percentage of cells with positive AVO staining was monitored using flow cytometry at 72 hours post treatment. Values shown are from a representative experiment with triplicate samples for each condition #p<0.05 compared to IR. ##p<0.05 compared to 1,25D3+IR
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S8/S9 Effects of pharmacologic autophagy inhibition on cell viability in IR and 1,25D3+IR treated cells ZR-75-1 cells were exposed to CQ, IR, 1,25D3+IR with and without CQ for 72 hours and cell viability was monitored by propidium iodide staining. Staining was quantified using flow cytometry. Positive staining is indicated by cells gated in the M1 region (S8). ZR-75-1 cells were exposed to CQ, IR+ CQ or 1,25D3+IR +CQ for 72 hours and apoptosis was monitored by annexin v/propidium iodide staining by flow cytometry (S9).
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