Fig. 3

LDL supplementation rescues C4-2B and PC3 cell growth in LPDS-containing media and is blocked by inhibition of ACAT. a Violin plots demonstrate that ACAT1 mRNA is overexpressed in prostate cancer tissue compared to normal in the TCGA dataset. b Violin plots demonstrate that ACAT1 protein level is increased in metastatic castrate-resistance prostate cancer (Met-CRPC) tissue compared to Benign prostate tissue. ACAT mRNA expression is associated with c shorter progression-free survival in TCGA prostate cancer dataset and d overall survival in Sboner et al. [57] dataset. e C4-2B cholesteryl ester levels and cellular protein following 24 h culturing in media supplemented with 10% FCS and increasing concentrations of the ACAT inhibitor Avasimibe. f C4-2B and g PC3 cholesteryl ester levels following 24 h culturing in media supplemented with 10% FCS, 10% LPDS, 10% LPDS plus 50 μg/ml of human LDL (LPDS+LDL), or 10% LPDS plus 50 μg/ml of human LDL and 1 μM of the ACAT inhibitor Avasimibe (LPDS+LDL+ACATi). h C4-2B cell proliferation cultured in media containing 10% FCS, 10% LPDS, LPDS+LDL, or LPDS+LDL+ACATi by IncuCyte and MTT. i PC3 cell proliferation cultured in media containing 10% FCS, 10% LPDS, LPDS+LDL, or LPDS+LDL+ACATi by IncuCyte and MTT. Data in a and b are represented as violin plots in GraphPad Prism: the horizontal line within the violin represents the median, and the dashed lines representing the quartiles. * P ≤ 0.05 vs. normal/benign; # P ≤ 0.05 vs. cancer by Mann-Whitney two-tailed t test (a) or one-way ANOVA (b) followed by Tukey’s multiple comparisons test. Data in c and d was analyzed using a two-sided log-rank test. Data in e and i are presented as mean ± SEM of at least three independent experiments performed in triplicate. * P ≤ 0.05 vs. FCS; $ P ≤ 0.05 vs. LPDS; † P ≤ 0.05 vs. LPDS+LDL by one-way ANOVA (e–g) or two-way ANOVA (h and i) followed by Tukey’s multiple comparisons test