Fig. 4: PROCR induces p65 activation via RSK1 to promote IL6 transcription.
From: PROCR diminishes the efficacy of radiation by impairing T-cell-mediated antitumour immunity
A Gene set enrichment analysis (GSEA) showing the top enriched pathways in parental (sgGFP) versus PROCR-knockout (sgPROCR) HONE1 cells. B RT-qPCR results for the mRNA expression of the indicated NF-κB-targeted genes in parental (sgGFP) and PROCR-knockout (sgPROCR) HONE1 cells. C Luciferase assay showing the NF-κB activity in parental (sgGFP) and PROCR-knockout (sgPROCR) HONE1 cells. D Western blot analysis of p65 in cytoplasmic (cyto) and nuclear (neu) fractions of parental (sgGFP), PROCR-knockout (sgPROCR), and PROCR-rescue (sgPROCR + PROCR-OE) HONE1 cells. Lamin B1 and α-tubulin were used as nuclear and cytoplasmic markers, respectively. E Western blot analysis of total and phosphorylated p65 in parental (sgGFP), PROCR-knockout (sgPROCR), PROCR-rescue (sgPROCR + PROCR-OE), and PROCR-OE HONE1 cells. F ChIP-PCR detection of the transcriptional regulation of p65 on IL6 in parental (sgGFP) and PROCR-knockout (sgPROCR) HONE1 cells. G Luciferase reporter assays showing the IL6 promoter activity in p65 or p65 plus PROCR overexpressed HONE1 cells. H Western blot analysis of total and phosphorylated RSK1, total and phosphorylated TBK1, and CDK6 in parental (sgGFP) and PROCR-knockout (sgPROCR) HONE1 cells. I Western blot analysis of total and phosphorylated RSK1, total and phosphorylated p65 in HONE1 cells pretreated with IgG or αPROCR. J Western blot analysis of total and phosphorylated p65 in parental (sgGFP), PROCR-knockout (sgPROCR), PROCR-rescue (sgPROCR + PROCR-OE), PROCR-rescue with RSK1-knockdown and PROCR-rescue with BI-D1870 (RSK1 inhibitor) in HONE1 cells. n = 3 independent experiments (B–J). One of three experiments is shown (D, E, H–J). Data are shown as mean ± SD (B, C, F, G). P values were calculated using two-tailed unpaired t test (C, F) or one-way ANOVA (B, G). Source data are provided as a Source Data file.
