Fig. 3

CPTH2 preferentially inhibits HAT-p300 in ccRCC 786-O cells. a In vitro HAT assay was performed on recombinant p300 (red), GCN5 (green), and PCAF (blue). Control (c) in presence of HAT inhibitor anacardic acid, AA (15 μM), and CPTH2 (400 and 600 μM). CPTH2 shows higher selectivity in the inhibition of p300. b Prolonged incubation with CPTH2 does not affect mRNA expression of p300. c treatment of ccRCC 786-O with CPTH2 (100 μM) at indicated times clears the immunostaining of p300 with respect to untreated and DMSO cells. d Silencing of p300 in 786-O cells caused decrease of p300-mRNA expression (light gray) and lowering of cell viability (gray) with respect to nc control. e Cytoskeleton of 786-O cells silenced for p300 (18 h) was stained with phalloidine. It is heavily modified and fully comparable to the cytoskeleton of cells treated with CPTH2 shown in Fig. 2a, and histogram with percentage of stress fibers/cell found after 24 h p300si (light gray) compared to control, nc (dark gray). f Histograms reporting the number of proliferating cells, percentage of stress fiber/cell and number of migrating cells in, respectively, nc control (black) and p300si cells (gray) grown in DMSO and in CPTH2 for 48 h