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. 2024 May 22;10:246. doi: 10.1038/s41420-024-02031-1

Fig. 4. Activated CREB signaling represses REST.

Fig. 4

Western blotting for REST, p-CREB1 (pS133, indicator of activation) and NE markers in prostate cancer cells upon treatments of CREB1 signaling activator 15 µM isoproterenol (ISO) or 10 µM Forskolin+ 0.5 mM IBMX for 24 h (A), or CREB1 signaling inhibitor ICI-118,551 (ICI, 10 µM) or synthetic peptide inhibitor of PKA (PKI, 10 µM) for 24 h (B). UT: untreated control, because ISO, ICI and PKI were dissolved in water. C Western blotting for REST, pS133-CREB1 and NE markers in PC3 cells carrying either an empty vector or CREB1-Y134F cDNA, a constitutively activated form of CREB1. D Western blotting for REST, pS133-CREB1, NE markers, EZH2 catalytic product H3K27me3 histone mark, loading controls histone 3 (H3) and beta actin in LNCaP cell-derived xenografts from NOD/SCID male mice, treated with saline or 10 mg/kg ISO for 54 days. E, F PC3-EV or PC3-REST cells were treated with DMSO control or CREB1 signaling activator combo 10 µM Forskolin (Fsk) + 0.5 mM IBMX for 24 h. mRNA levels of indicted genes were normalized to GAPDH control. F Morphology of PC3-EV and PC3-REST cells treated with DMSO or Fsk+IBMX.