Fig. 1: BMAL1 is elevated and active in ccRCC.

A Phylogenetic tree for bHLH-PAS proteins. B Percent sequence identity for bHLH and PAS domains in BMAL1 and ARNT. C Detection of BMAL1 (transcripts per million, TPM) in RNA sequencing data from tumors and adjacent normal tissues in cancer genome atlas projects: colorectal adenocarcinoma (COAD, n = 82 non-tumor, n = 962 tumor), lung adenocarcinoma (LUAD, n = 59 non-tumor, n = 539 tumor), breast cancer (BRCA, n = 113 non-tumor, n = 1111 tumor), kidney clear cell renal cell carcinoma (KIRC, n = 72 non-tumor, n = 541 tumor), and renal papillary carcinoma (KIRP, n = 100 non-tumor, n = 872 tumor). D Nuclear BMAL1 protein detected in human kidney biopsy samples from ccRCC (n = 138), other renal cancers (n = 42), and non-tumor kidney tissue (n = 30). E Clock correlation distance (CCD) heatmaps calculated from RNA sequencing data from tumors and adjacent normal tissues in the Cancer Genome Atlas projects. In (E) one-sided p-values for non-tumor vs. tumor samples are calculated from permutation testing as described in detail in ref. ¹. F Luminescence detected in cells expressing destabilized luciferase under the control of the PER2 promoter in 786O, A498, or RCC4 cell lines in which circadian rhythms were synchronized by treatment with dexamethasone (red) or horse serum (orange). G Quantitation of the rhythmic amplitude for data as in (F) for 786 O (n = 3 biological replicates per condition) or RCC4 (n = 4 biological replicates per condition) cells expressing Per2-Luciferase with (green) or without (black) concomitant expression of VHL and synchronized with dexamethasone. Error bars represent s.d. P values calculated by two-sided t-tests. H Detection of the indicated proteins by immunoblot in cell lysates prepared from 786O cells at the indicated times after treatment with dexamethasone or without synchronization (ns) or expressing VHL. Data represent one of two independent experiments with similar results. The samples derive from the same experiment but one gel for BMAL1, HIF2α, and ACTIN, and another for ARNT and ACTIN were processed in parallel. In (C), boxplots depict the median and interquartile range (IQR), whiskers extend either to the minimum or maximum data point or 1.5 × IQR beyond the box, whichever is shorter. Outliers (values beyond the whisker) are shown as dots. In (C,D) **P = 0.00149, ****P < 0.0001 by two-way ANOVA. Source data are provided as a Source Data file.