Supplementary Methods, Figures 1 - 6, Tables 1 - 2 from Prostate Tumor Growth Is Impaired by CtBP1 Depletion in High-Fat Diet–Fed Mice
Authors: Moiola Cristian P. , De Luca Paola , Zalazar Florencia , Cotignola Javier , Rodríguez-Seguí Santiago A. , Gardner Kevin , Meiss Roberto , Vallecorsa Pablo , Pignataro Omar , Mazza Osvaldo , Vazquez Elba S. , De Siervi Adriana 
Published: April 2023

<p>PDF file - 556K, Table S1: Genesets enriched in PC3.pGIPZ xenografts. Table S2: Genesets enriched in PC3.shCtBP1 xenografts. Supplemental Figure 1: (A) Crystal violet stained NIH3T3 cells foci transfected with pcDNA3.CtBP1 or beta-gal expression vectors. (B) CtBP1 RT-qPCR from PC3.CtBP1, PC3.shCtBP1 and PC3.pGIPZ stable cells using specific CtBP1 and Actin beta primers. Fold induction was calculated normalizing data to Actin beta and control. Bars represent the average and standard deviation of one representative experiment.***p < 0.001. Supplemental Figure 2: Box plots for (A) triglycerides, (B) glycemia and (C) estradiol serum levels for CD or HFD fed mice were shown. Boxes represent the interquartile range, the horizontal line within each box represents the median, and the upper and lower whiskers represent the standard deviation of one independent experiment. (D-E) Representative photograph of H&E staining from kidney and liver of mice fed with CD or HFD (Magnification 400x & 250x respectively). Supplemental Figure 3: (A) Testosterone and (B) estradiol levels from CD or HFD xenografts mice. Plotted boxes represent the interquartile range, the horizontal line within each of the boxes represents the median, and the upper and lower whiskers represent the standard deviation. * p < 0.05 (C) Specific CtBP1 RT-qPCR from tumors of CD or HFD fed mice. Fold induction was calculated normalizing data to Actin beta and control. Bars represent the average and standard deviation from 3 independent mice. * p < 0.05. Supplemental Figure 4: (A) GSEA from microarray data from HFD tumor xenografts. enrichment plots from genesets enriched in PC3.shCtBP1 xenografts. We showed the most significant over-representive cell adhesion categories selected from GSEA analysis from microarray data. (B) CDH1 RT-qPCR from the HFD fed mice tumors was performed. Fold induction was calculated normalizing data to Actin beta and control. Bars represent the average and standard deviation from 3 mice. * p < 0.05 (C) GSEA from microarray data from HFD tumor xenografts. The most significant enrichment plots for hormone pathways genesets in PC3.shCtBP1 xenografts were shown. Supplemental Figure 5: (A) Enrichment plots from genesets enriched in PC3.pGIPZ xenografts. Enrichment plots for the most significant over-representive categories selected from GSEA analysis from microarray data were shown. Listed above we found enrichment plots associated to cell cycle regulation processes and proliferation. Supplemental Figure 6: (A-B) BRCA1 RT-qPCR from (A) 22Rv1 or (B) LNCaP cells grew in charcoal FBS-medium and exposed to testosterone or vehicle for 24h. Fold induction was calculated normalizing data to Actin beta and control. (C-D) BRCA1 RT-qPCR from (C) 22Rv1 or (D) LNCaP cells, pre-treated for an hour with letrozole, were then grown as indicated before. Fold induction was calculated normalizing data to Actin beta and control. Data represents the average and standard deviation of one representative experiment. * p < 0.05.</p>