Figure 5.

Validation of RNA-Seq data from AtT20 cells treated with JQ1- or JQ1. Transcription of the apoptosis associated genes Capn9, Dffb, Birc3 and Nfkb1 was examined using qRT-PCR, after 48 h JQ1 treatment (A). Expression of cIAP2 (encoded by Birc3) and Nfkb1 after JQ1 treatment was confirmed by Western blot analysis (B) and was significantly decreased compared to control treated cells, as indicated by densitometry analysis (C). Transcription of the SSTR2 proliferation pathway associated genes Sstr2, Npr1, Gnb4, Gng4, Gnb3 and Nos1 was examined using qRT-PCR, after JQ1 treatment (D). The expression of SSTR2 after JQ1 treatment was confirmed using Western blot analysis (E) and was significantly downregulated, compared to control treated cells, as indicated by densitometry analysis (F). For all experiments untreated (UT), vehicle only (DMSO) and JQ1- were used as negative controls, and Gapdh (a housekeeper gene) was used as a loading control. All qRT-PCR experiments were performed in n = 4 technical and n = 4 biological replicates, and Western blots performed in n = 5 biological replicates. Statistical significance is relative to DMSO vehicle only treatment with * P < 0.05, ** P < 0.005 and *** P < 0.0005.