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. 2016 Sep 30;291(47):24538–24550. doi: 10.1074/jbc.M116.746370

FIGURE 6.

FIGURE 6.

ZBTB33 regulates the RB1-E2F pathway. A, representative immunoblots for whole (pan) and phosphorylated RB1 in HeLa and HEK293 cells (100 and 12 μg of total protein per lane, respectively) after ZBTB33 depletion. Note, HeLa and HEK293 β-actin immunoblots were analyzed using different exposure intensities. B and C, 6×E2F-luciferase reporter was transfected into control and ZBTB33-depleted HeLa and HEK293 cells, and luciferase activity was quantitated. Luciferase reads, normalized to Renilla activity, represent endogenous E2F activity. D, immunoblot analysis for ZBTB33 in HeLa cells transfected with ZBTB33 overexpression plasmids under control of either a 2-kb minimal endogenous ZBTB33 promoter or a highly active CMV promoter. E and F, 6×E2F-luciferase reporter was transfected into mock control or ZBTB33-overexpressing cells demonstrating dose-dependent E2F activity in HeLa cells and a decreased E2F activity in HEK293 cells. G, representative immunoblots for E2F1–5 proteins in HeLa and HEK293 cells after ZBTB33 depletion. *, p < 0.05; **, p < 0.01; ***, p < 0.005 by Student's t test.