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. 2017 Aug 17;9(2):1602–1616. doi: 10.18632/oncotarget.20308

Figure 5. Inhibition of autophagy increases the sensitivity of LM7 osteosarcoma cells to gemcitabine (GCB)-induced cytotoxicity.

Figure 5

A LM7 cells were pretreated for 30 minutes with 20μM hydroxychloroquine (HCQ), 1μM GCB, or both for 48 hours. Cell viability was measured using Trypan blue exclusion assay. B Apoptosis was measured using propidium iodide/Annexin V staining. Cells were treated with GCB alone, HCQ alone, or HCQ combined with GCB for 48 hours. After incubation with Annexin V-FITC in a buffer containing propidium iodide, cells were analyzed using flow cytometry. C. After knockdown of BECN by shRNA, whole cell lysates were analyzed for BECN expression by Western blot. β-actin was used as a loading control. D. shRNA-BECN cells and LM7 control cells were treated with 1μM GCB for 48 hours. Cell viability was measured using Trypan blue exclusion assay. Means ± standard deviation of three independent experiments are shown. *p < 0.05 compared with control.