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. 2018 Dec 6;160(2):276–291. doi: 10.1210/en.2018-00920

Figure 2.

Figure 2.

Intracellular signaling to POMC expression in acidic pH. (A) Representative Western blots for Pomc, Crhr1, Nur77, Tpit, Pitx1, Neurod1, phospho-Erk1/2 (p-Erk1/2), Erk1/2, phospho-CamkII (p-CamkII), CamkII, phospho-Creb (p-Creb), Creb, and β-actin in AtT-20 cells after 24-h treatment in pH-adjusted medium. (B) Quantification of each protein level normalized to β-actin in (A). (C) Western blots for p-CamkII, CamkII, Tpit, Pomc, and β-actin in AtT-20 cells after 24-h treatment at pH 7.4 or pH 6.8 with CaMKII inhibitor KN-62. (D) Quantification of each protein level normalized to β-actin in (C). (E) Western blots for phospho-Creb (p-Creb), Creb, Pomc, and β-actin after 24-h treatment in pH 7.4 and pH 6.8 medium with CREB inhibitor H-89. (F) Quantification of each protein level normalized to β-actin in (E). (G) Structures of partially deleted or point mutated luciferase reporter plasmids of the rPomc promoter (–480/+63, –379/+63, –323/+63, –200/+63, –155/+63, –100/+63, –34/+63) in Tpit, Pitx1, Tpit/Pitx1, and Neurod1 (left) and relative luciferase activity normalized to values in AtT-20 cells transfected with pGL4 (right). Results are mean ± SD. *P < 0.05 vs pH 7.4; **P < 0.01 vs pH 7.4. n.s., not significant.