Fig 1: ORP4L is required for C33A, HeLa and CaSki cells proliferationA. ORP4/OSBP2 gene transcripts in HeLa/C33A/CaSki cells were detected by RT-PCR. Actin expression was used as an internal control. B. HeLa cells were transfected with 60 pmol siORP4L.1, siORP4L.2 or siNT in 12-well plates. Silencing efficiency was accessed at mRNA (qRT-PCR analysis, left panel) and protein level (western blots analysis, right panel). C. ORP4L/M/S mRNA was measured by semi-quantitative RT-PCR using cDNAs prepared from HeLa cells transfected with siORP4L.1, siORP4L.2 or siNT. D. C33A cells were transfected with siORP4L.1, siORP4L.2 or siNT, flow cytometry analysis of cell proliferation was conducted after 0 hr, 24 hr, 48 hr, and 72 hr with CFSE staining. E. HeLa (left) and CaSki (right) cell proliferation upon ORP4L knockdown analyzed by flow cytometry and CCK-8, respectively. F. Analysis of C33A cells proliferation after transfection with ORP4L cDNA or empty vector; flow cytometry analysis of cell proliferation was carried out after 0 hr, 24 hr, 48 hr, and 72 hr with CFSE staining. G. HeLa (left) and CaSki (right) cell proliferation upon ORP4L knockdown analyzed by flow cytometry and CCK-8, respectively. The data represent mean ± S.D. from three individual experiments (n = 3, *p < 0.05, **p < 0.01).