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. 2013 Apr 15;123(5):2103–2118. doi: 10.1172/JCI61820

Figure 2. Inhibition of glioma cell growth in vitro by knockdown of PCBP2.

Figure 2

(A) Western blot of PCBP2 expression in 3 glioma cell lines (T98G, U87MG, and U251) transiently transfected with the control siRNA or PCBP2 siRNA for 48 to 72 hours using Lipofectamine 2000. β-Actin was used as a loading control. (B) MTT assay on the same 3 glioma cell lines after transfection with the control siRNA or PCBP2 siRNA as above. Data are presented as the mean ± SD and are representative of 3 wells. *P < 0.05 compared with the control siRNA by a 2-tailed Student’s t test. (C) Approximately 72 hours after transfection, the 3 glioma cell lines were analyzed by flow cytometry. The proportions of cells in the G1, G2, and S phases of the cell cycle are depicted in the histograms. *P < 0.05 compared with the control siRNA by a 2-tailed Student’s t test. (D and E) Representative Western blot showing P27, P21, and P16 protein levels (D) and expression levels of differentially phosphorylated pRb (E) in the transfected glioma cell lines. P16 was deleted in U87MG and U251 cells. (F) Nuclear TUNEL staining for apoptotic cells in the glioma cell lines after approximately 72 hours of transfection. The ratio of TUNEL-positive cells was calculated (n = 5) and plotted on the histogram. *P < 0.05 compared with the control siRNA by a 2-tailed Student’s t test. (G) Representative Western blot showing (cleaved) caspase-3 and its substrate (cleaved) poly (ADP-ribose) polymerase (PARP) in the transfected glioma cell lines.