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. Author manuscript; available in PMC: 2011 Mar 15.
Published in final edited form as: Cancer Res. 2010 Jan 19;70(3):1247–1255. doi: 10.1158/0008-5472.CAN-09-2774

Figure 2. Role of PI-PLC γ2 in TDCA-induced NOX5-S expression.

Figure 2

(A) A typical example of Western blot analysis and summarized data showed that transfection with PI-PLC γ2 siRNA significantly decreased PI-PLC γ2 protein expression in FLO EA cells, indicating that PI-PLC γ2 siRNA effectively knocked down PI-PLC γ2 protein expression (n=3). (B) A typical example of Western blot analysis and summarized data show that knockdown of PI-PLC γ2 protein significantly decreased TDCA-induced NOX5-S expression. (C) Knockdown of PI-PLC γ2 protein significantly decreased TDCA-induced H2O2 production. (D) Knockdown of PI-PLC γ2 protein significantly decreased TDCA-induced thymidine incorporation both in FLO and OE33 cells. The data suggest that PI-PLC γ2 protein may contribute to TDCA-induced NOX5-S expression, H2O2 production and cell proliferation in EA cells. Transfection of siRNA was carried out with Lipofectamine 2000. Per well, 75 pmol of siRNA duplex of PI-PLC γ2 or control siRNA formulated into liposomes were applied. After a 4-h transfection, the transfection medium was replaced with regular medium. 24 h after transfection, cells were treated with low dose of TDCA for 24 h, and then culture medium and cells were collected for measurements. N=3, ANOVA, * P<0.02, compared with control group and control siRNA group; ** P<0.02, compared with control siRNA group and PI-PLC γ2 siRNA group; *** P<0.001, compared with control siRNA+ TDCA group, ▲ P<0.001, compared with control siRNA+ TDCA group.