(A) RNA-seq analysis indicates the genes commonly upregulated by RB1 knockdown and palbociclib treatment in PC-3 cells.
(B) Ingenuity pathway analysis (IPA) of common targets shown in (A) (ranked by p-values).
(C) Heatmap showing a subset of RB knockdown and palbociclib-coregulated genes involved in T cell regulation and TNF-α-NF-κB pathways.
(D) Gene set enrichment analysis (GSEA) of the common targets shown in (A).
(E) UCSC Genome Browser screen shot of RNA-seq tracks for T cell/NF-κB pathway genes (PD-L1, GADD45B, NR4A2, and CD83) commonly upregulated by RB knockdown and palbociclib treatment.
(F and G) PC-3 cells treated with or without palbociclib (5 μM) for 24 hr (left) or infected with lentivirus expressing control or RB-specific shRNAs (right) were harvested for RT-qPCR (F) and ChIP-qPCR (G) analysis of p65 binding at the promotor of PD-L1, GADD45B, NR4A2, and CD83 genes. All data are shown as mean values ± SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.