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. 2019 Aug 2;11(8):1105. doi: 10.3390/cancers11081105

Figure 6.

Figure 6

CREB-binding protein (CBP) is linked to the gene expression regulatory role of High Mobility Group A1. (A, B, and C—left side) Western blot (WB) analyses performed with MDA-MB–231 cell lysates. Cells were grown with complete medium (lane 1) or serum-starved medium (lanes 2, 3 and 4) for 24 h before treatment with 20 μM C646 or 0.2% dimethyl sulfoxide (DMSO) as a control (CTRL). UT (untreated) represents cells treated neither with C646 nor with DMSO (A—left side). MDA-MB–231 cell lysates silenced for CBP and p300 (siCBP and sip300) and their relative controls (siCTRL) (B and C—left side, respectively). All WB experiments were performed in biological triplicate, providing consistent results. Representative images are shown. (A, B, and C, right side) qRT-PCR analyses of serum-starved MDA-MB-231 cells treated for 24 h with 20 μM C646 (A, right side) MDA-MB-231 cells silenced for CBP (B, right side) or p300 (C, right side) and their respective controls (DMSO and siCTRL). The four HMGA–signature genes (Hsss) were analysed together with CBP and p300. Glyceraldehydes-3-phosphate dehydrogenase (GAPDH) or cyclophilin-33 (CYP33) were used as internal reference genes. Mean, standard deviations (n = 3), and statistical significance (t-test) are indicated (*: p < 0.05; **: p < 0.01; ***: p < 0.001).