Fig. 4: KLHL42 is a transcriptional target of MYC that mediates CDK4/6i resistance.
From: MYC induces CDK4/6 inhibitors resistance by promoting pRB1 degradation
a Screenshot of the UCSC genome browser showing ChIP-seq signal profiles of MYC in the KLHL42 gene locus in different human cell lines, as previously reported55,56. b ChIP‒qPCR analysis of MYC binding at the promotor of the KLHL42 gene in T24 and UMUC14 cells. Data were shown as the mean ± SD of three independent experiments (n = 3). Two-tailed unpaired Student’s t-test. P values based on the order of appearance: 0.0001, and 0.0004. Control or MYC-knockdown T24 and UMUC14 cells were harvested for western blotting (c) and RT‒qPCR analyses (d). In d, data were shown as the mean ± SD of three independent experiments (n = 3). Two-tailed unpaired Student’s t-test. P values based on the order of appearance: 0.0025, 0.0016, 0.0003, and 0.0004. e, f Representative images of IHC analysis with anti-RB1, anti-Myc and anti-KLHL42 antibodies on FFPE samples of prostate specimens from Pbsn-Cre and High Myc transgenic mice and the quantitative data of Rb1, Myc and Klhl42 staining were shown in f. Scale bar in 10 X fields: 200 μm; Scale bar in 40 X fields: 20 μm. In f, data were shown as the mean ± SD (Pbsn-cre n = 5, Hi-Myc tumor n = 8). Two-tailed unpaired Student’s t-test. P values based on the order of appearance: 0.0017, 2.66E-05, and 1.17E−06. g T24 and UMUC14 cells infected with the indicated lentivirus expressing shRNAs were harvested for western blotting. T24 and UMUC14 cells infected with the indicated lentivirus expressing shRNAs treated with vehicle or palbociclib were harvested for colony formation assay (h). In i, data were shown as the mean ± SD of three independent experiments (n = 3). Two-tailed unpaired Student’s t-test. P values based on the order of appearance: 0.0018, 0.0009, 0.0006 and 0.0004; 0.9882, 0.0002, 0.0003 and 0.0012. T24 (j) and UMUC14 (k) cells infected with the indicated lentivirus expressing shRNAs treated with palbociclib were harvested for the cell viability assay. Data were shown as the mean ± SEM of three independent experiments (n = 3). Source data are provided in this paper. Similar results for c and g panels were obtained in three independent experiments.
