Figure 5.

Overexpression of PRDM1 induces cell cycle arrest and apoptosis in LCL cells. CCL159 and JY25 cells were transfected either with pMSCV-PIG vector or pMSCV-PRDM1 plasmid. Total RNA or protein was collected at 48 h time point, PRDM1 expression was determined by real-time PCR (a) or western blotting (b). Lamin B was included as a loading control. The lower band seen (indicated by asterisk) in (b) is most likely a non-specific band, as previously reported.⁴⁸ (c) Cells collected at 48 h were stained with Annexin V and 7AAD using apoptosis detection kit (BD Biosciences, San Jose, CA, USA) and subjected to flow cytometry analysis. (d) Cell cycle analysis of GFP positive cells at indicated time point. The percentage of each cell cycle phase distribution in CCL159 (e) and JY25 (f) cells was calculated by FlowJo software (Ashland, OR, USA) and graph was generated using the prism 6 software. Error bars represent s.e.m. from technical replicates. *P<0.05; **P<0.01; ***P<0.005. GFP, green fluorescent protein.