Fig. 9: PKCλ/ι loss increases selective translation to promote a TGFβ resistant program. | Nature Communications

Fig. 9: PKCλ/ι loss increases selective translation to promote a TGFβ resistant program.

From: Increased translation driven by non-canonical EZH2 creates a synthetic vulnerability in enzalutamide-resistant prostate cancer

Fig. 9

a Schematic of monosome and polysome isolation by sucrose gradient fractionation. b Polysome profiles of sgPRKCI and sgC LNCaP cells treated with 10 μM of ENZA for 72 h (n = 3 biological replicates). c Top 5 Hallmark pathways enriched in translationally efficient mRNAs upregulated in LNCaP sgPRKCI, by HOMER software (n = 3 biological replicates). d Ingenuity Pathway Analysis for translationally efficient mRNAs of LNCaP sgPRKCI vs sgC determined by Xtail (n = 3 biological replicates). e Volcano plot of translationally efficient mRNAs of LNCaP sgPRKCI vs sgC determined by Xtail (n = 3 biological replicates) (Blue= neuronal genes, red= EMT-related genes). f Immunoblots in sgPRKCI and sgC LNCaP cells transduced with the indicated siRNAs, treated as in (a) and quantification (n = 3 independent experiments). g Upstream regulator analysis of translationally efficient genes enriched mRNAs in sgPRKCI, treated with ENZA for 72 h (n = 3 biological replicates). h Dose-response curves to determine the IC50 of ENZA either treated with vehicle or 20 μM galunisertib (Gal) in sgC and sgPRKCI LNCaP cells for 6 days. IC50 value is the average of two biological replicates. i Growth curves of sgC and sgPRKCI LNCaP cells treated with 10 μM of ENZA and 20 μM of galunisertib alone or combined. Representative experiment of two biological replicates. j Growth curves of PRKCI-overexpressing (PRKCI-OE) WCM1078 organoids treated as in (i). Representative experiment of three biological replicates. k PKCλ/ι’s dual role in EZH2 regulation. First, by controlling its stability, mediating its interaction with RBBP6. Second, by facilitating the transition of EZH2 from a Polycomb repressor to a transcriptional coactivator of YY1. This transition mediates resistance to enzalutamide induced by the loss of PKCλ/ι. Data shown as mean ± SEM of 3 biological replicates (f), mean ± SD of technical triplicates (i), mean ± SD of 3 biological replicates (j). Two-way ANOVA (i, j). Two-tailed unpaired Student’s t-test (f). Source data are provided as a Source Data file.

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