Fig. 1: PPARG expression is necessary for tumour development in vivo.

A Immunoblot of PC3-M CRISPR knockdown of PPARG clones (KO1, 2 & 8) and Scrambled control (Scrm), representative image of three experiments. Values above each band indicate densitometry value of band as normalised to Actin loading control and compared to first band (NTS), N.D not detected. B qPCR analysis for PPARG levels in clones KO1, 2 & 8, and control Scrm, three independent experiments and three technical replicates per experiment, error bars show SEM. C Ultrasound measurement of tumour size (KO clones & Scrm control) over time in days, ‘*’ denotes statistical significance p ≤ 0.05 as determined by 2-way Anova and Dunnett’s multiple comparison test. D Final tumour weight (KO clones & Scrm control) upon necropsy, ‘*’ denotes statistical significance p ≤ 0.05 as determined by one-way Anova and Holm-Sidaks multiple comparison test. For both C and D graphs represent 23 mice, at least 5 mice per cell line; ultrasound performed every two weeks until endpoint. All mice taken at the same time when the first mouse reached endpoint. E IHC images of PPARG KO derived orthograft tumours. Representative images from PPARG KO tumours stained for PPARG and FASN, scale bar shows 100 μm. F Immunoblot of DU145 PPARG over-expressing clones (OE12, 18 & 19) and control (EV7), representative image of three independent experiments. Values above each band indicate densitometry value of band as normalised to Actin loading control and compared to first band (EV7), N.D not detected. G qPCR analysis for PPARG levels in OE12, 18 & 19 and control EV7 qPCR three independent experiments and three technical replicates per experiment, error bars show SEM. H Ultrasound measurement of tumour size over days in OE clones & EV7 control, statistical significance is denoted by * where p ≤ 0.05 as determined by 2-way Anova and Dunnetts multiple comparison test. I Final tumour weight upon necropsy in OE clones & EV7 control. Statistical significance is denoted by * where p ≤ 0.05 determined by one-way Anova and Holm-Sidak’s multiple comparisons test. For H and I graphs represent 24 mice, 6 mice per cell line, ultrasound performed every two weeks until endpoint. All mice were taken at the same time when the first mouse reached endpoint. J IHC images of PPARG OE derived orthograft tumours. Representative images from tumours stained for PPARG and FASN, scale bar shows 100 μm. K IHC image analysis for PPARG and FASN. Graph displays average percentage positive cells over at least three samples per clone with error bars giving SEM. Statistical significance, where found, denoted by * with p ≤ 0.05 determined by Mann–Whitney.