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. 2010 Apr 27;285(25):19106–19115. doi: 10.1074/jbc.M109.099770

FIGURE 5.

FIGURE 5.

KLF2 negatively regulates FABP5, and the two proteins are inversely regulated by HRG-β1 and NF-κB. a–c, MCF-7 cells were infected with adenoviruses harboring GFP or GFP-KLF2 for 48 h. Expression level of mRNA for KLF2 (a) or FABP5 (b) were measured by Q-PCR. Data are the mean ± S.D. (n = 3). *, p < 0.025. c, cells were lysed 24 or 48 h after transfection, and lysates were analyzed by immunoblots using denoted antibodies. d, MCF-7 cells were transfected with si-control or vector encoding Si-KLF2 (Santa Cruz Biotechnology, Inc.), and cells were treated with HRG-β1 (30 ng/ml, 6 h). Expression levels of FABP5 and the internal control cyclophilin were measured by quantitative PCR (FABP5, forward (gcagacccctctctgcac) and reverse (tcgcaaagctattcccactc); cyclophilin, forward (cctaaagcatacgggtcctg) and reverse (tttcactttgccaaacacca)). *, p = 0.0004 versus non-treated cells transfected with si-control; **, p = 0.0003 versus non-treated cells transfected with si-control; ***, p = 0.01 versus HRG-β-treated cells transfected with si-control. e, MCF-7 cells were infected with control adenovirus (Ad-GFP) or adenovirus overexpressing KLF2 (Ad-KLF2) and treated with HRG-β1 for 24 h. Cell proliferation was assessed by MTT assays. *, p = 0.001 versus non-treated cells infected with Ad-GFP; **, p < 0.001 versus si-control-infected untreated cells; ***, p < 0.001 versus Ad-GFP-infected, HRG-β1-treated cells. f, serum-starved MCF-7 cells were treated with the NF-κB inhibitor PDTC (100 μm) for 1 h before the addition of HRG-β1 (30 ng/ml, 4 h). KLF2 mRNA levels were measured by Q-PCR. Data are the mean ± S.D. (n = 3). p = 0.009 (*) and p = 0.05 (**) versus non-treated control. g, MCF-7 cells were transfected with a control vector or a vector harboring dominant negative IκB-α (SRIκB) for 36 h. ev, empty vector. Cells were serum-starved overnight and treated with HRG-β1 for 4 h. FABP5 mRNA levels were measured by Q-PCR. Data are the mean ± S.D. (n = 3). p = 0.015 (*) and p = 0.02 (**) versus empty vector-transfected non-treated empty vector control.