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. 2014 Sep 26;5(21):10692–10708. doi: 10.18632/oncotarget.2530

Figure 4. Administration of hUCESCs-CM to MDA-MB-231 cells delayed the cell cycle and increased apoptosis.

Figure 4

(A): MDA-MB-231 cells were treated for 48 h with DMEM plus 10% FBS (+FBS), incomplete medium (DMEM without FBS, -FBS), or 48-h hUCESCs-CM, and then subject to flow cytometry using propidium iodide (PI). Percentage of cells (mean + standard deviation) in each phase is shown. (B): Western blot of cyclin A, cyclin B, cyclin E, cyclin D1, and GAPDH (used as loading control) of protein extracts from MDA-MB-231 cells treated for 48 h as described in (A). (C): Apoptosis was determined in MDA-MB-231 cells cultured for 48 h with complete (+FBS), incomplete (−FBS), or hUCESCs-CM by flow cytometry using Annexin V/PI. Annexin V+/PI- and Annexin V+/PI+ indicated early and late apoptosis, respectively. (D): Western blot of Caspase 8, -12, -9, activated caspase 3, and cleaved PARP of MDA-MB-231 protein extracts as indicated in (C). (E): Western blots of the anti-apoptotic Bid, cleaved Bid, and Bim proteins in MDA-MB-231 extracts treated as in (C). GAPDH was used as loading control.