Figure 5.

p300 downregulation decreases cell migration and invasion. (A) Wound scratch assays were performed on confluent layers of PC3-DR shCtrl, shp300-1, and shp300-2 treated with 100 ng/mL doxycycline. Images were taken 48 h after scratch and analyzed by ImageJ (n = 6). (B) PC3-DR were seeded in Boyden chambers and shp300 sequences were activated with 100 ng/mL doxycycline for 72 h. Cell migration was measured after staining with calcein and visualized by fluorescence microscopy (n = 4). (C) Invasion assays were conducted as in B, except that Boyden chambers were pre-coated with Matrigel (n = 3). Values indicated in A–C denote mean + s.e.m. (one-way ANOVA). (D) Wound scratch assays on PC3-DR treated with 10 µM CPI-637. Data represent mean + s.e.m. (t-Test, n = 3). (E) Migration (n = 4) and (F) invasion assays (n = 5) of PC3-DR treated with 20 µM CPI-637. Values indicated are mean + s.e.m. (t-test). (G) Immunofluorescence staining for vimentin (red) and quantification relative to counterstaining of nuclei (blue). Original magnification 630×. (H) mRNA (qPCR) and protein expression (Western blot) of vimentin. Data in G-H represent mean + s.e.m. (one-way ANOVA, n = 5).