Figure 6.

POMC expression and ACTH secretion after JQ1 treatment. QRT-PCR analysis confirmed the significant down-regulation of Pomc transcription after 48 h JQ1 treatment (A). Down-regulation of POMC protein expression was also confirmed by Western blot analyses at 24 h, 48 h and 72 h after JQ1 treatment, compared to JQ1- treated cells; POMC is observed as two bands corresponding to pre-pro-POMC and pro-POMC (calnexin was used as a loading control) (B). Densitometry analysis confirmed that POMC expression was significantly down-regulated at 24 h, 48 h and 72 h (C). Concentrations of ACTH in the media from the same cells, which reflected ACTH secretion by these cells, significantly decreased at 48 h and 72 h after treatment with JQ1, when compared to JQ1- treated cells (D). DMSO or JQ1-treated cells were used as negative controls, with qRT-PCR experiments performed in n = 4 technical and n = 4 biological replicates, and Western blot and ELISAs performed in n = 4 biological replicates. Statistical significance is relative to JQ1- treatment with *P < 0.05, **P < 0.005 and ***P < 0.0005.