Extended Data Fig. 1: Genes encoding F-box proteins are not highly mutated in human cancers.
a, Heat map showing percentages of samples mutated in the indicated genes, across individual tumor types or pan-cancer, and ranked by their percent mutation rate in pan-cancer. Genes encoding F-box proteins are in black; different cancer genes are in blue. b, Scheme showing the CpG island within the FBXL7 promoter and the primers used to assess FBXL7 promoter methylation by methylation-specific PCR analysis. c, Total RNA extracts from the indicated pancreatic cells were analyzed for FBXL7 mRNA levels by qPCR. Mean ± s.d. is shown; n = 3 independent experiments; P values are from unpaired, two-tailed t-test. d, Total DNA extracts from H6c7, AsPC1, and COLO357 pancreatic cells were subjected to bisulfite modification and sequencing of a FBXL7 promoter region within the CpG island. The table shows the percentage of methylation of the FBXL7 promoter. e, Total mRNA extracts from the indicated prostate cells were analyzed for FBXL7 mRNA levels by qPCR. Mean ± s.d. is shown; n = 3 independent experiments; P values are from unpaired, two-tailed t-test. f, Total DNA extracts from PNT1A, LAPC4, PC-3, PC-3M, and PC-3M-LN4 prostate cells were subjected to bisulfite modification and sequencing of a FBXL7 promoter region within the CpG island. The table shows the percentage of methylation of the FBXL7 promoter. g, Co-expression of FBXL7 and AR mRNAs was analyzed in the prostate TCGA dataset (cBioPortal; n = 450). Two-tailed non-parametric Spearman correlation was used. The linear regression line is shown in red. h, Total mRNA extracts from AsPC1, PC-3, PC-3M and PC-3M-LN4 cells were analyzed for FBXL7 mRNA levels by qPCR upon addition of decitabine for the indicated times. Mean ± s.d. is shown; n = 3 independent experiments; P values are from unpaired, two-tailed t-test. i, PL45, AsPC1, PNT1A, LNCaP, and PC-3 cells were transfected with two different siRNAs to FBXL7 (each individually) or a non-targeting (NT) siRNA. Twenty-four hours after transfection, cells were harvested and plated in 96-well plates in triplicates. Cell proliferation was assessed at the indicated times by measuring absorbance (OD) at 590–650 nm. A representative experiment out of two, each performed in triplicate, is shown. Mean ± s.d. is shown.
