Figure 3
From: Silencing of Kruppel-like factor 2 by the histone methyltransferase EZH2 in human cancer
KLF2 induces apoptosis, directly activates the cell cycle-inhibiting genes p15/CDKN2B and p21/CDKN1A and inhibits cell proliferation. (A) Enhanced Annexin V expression relative to empty vector-transfected cells upon KLF2 transfection. The right panels are the original red color/FL-4 (annexin-Cy5) histograms that show how KFL2 transfection induces apoptosis, demonstrated by the increase in the number of cells that incorporate higher amounts of annexin V. (B) U2OS cells transfected with a FLAG-tagged KLF2 (pCMV-Tag2B-KLF2) expression vector show increased expression of p15Ink4b and p21CDKN1A, determined by qRT–PCR (a) and western blot (b). (C) The ectopically expressed KLF2 (FLAG-tagged KLF2) occupied the p15Ink4b and p21CDKN1A promoters in association with a gain of RNAP-S2 occupancy and the H3-K4 trimethylation mark determined by qChIP. (D) MTT (left) and colony formation (right) assays reveal that stable KLF2-expressing U2OS cells grow more slowly than cells transfected with control vector. (E) Enhanced proliferation of normal MRC5 fibroblasts after transfection of shRNA against KLF2 in comparison with scrambled shRNA. P-values obtained from Student’s t-test.
