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. 2021 May 10;118(20):e2023871118. doi: 10.1073/pnas.2023871118

Fig. 2.

Fig. 2.

MAPK signaling involved in acidosis-associated gene transcription. (A) Western blot depicting phosphorylation of ERK, MSK, and CREB at pH 7.4 or exposed to pH 6.0 for 15, 30, and 60 min. (B) Densitometry analysis and representative Western blot of phospho-CREB and pan-CREB in T84 IEC pretreated with 10 μm MSK1 inhibitor following 30 min exposure to pH 7.4 and 6.0 HBSS+ (n = 3). (C) FosB expression in T84 IEC pretreated with 10 μm MSK or MEK inhibitors following 6 h exposure to pH 7.4 and 6.0 HBSS+ (n = 3). (D) Densitometry analysis and representative Western blot of MSK1 expression in shRNA lentiviral transduced T84 IEC (n = 3). (E) Representative Western blot and densitometry analysis of phospho-CREB and pan-CREB in shCont and MSK1 KD IEC following 30 min exposure to pH 7.4 and 6.0 HBSS+ (n = 3). (F) FosB, IL-8, and TNF-α expression following 6 h exposure to pH 7.4 and 6.0 HBSS+ in shCont and MSK1 KD IEC (n = 3). (G and H) Analysis of secreted TNF (G) and IL-8 (H) following 16 h exposure to pH 7.4 and 6.0 HBSS+ in shCont and MSK1 KD IEC (n = 3). Data from each experiment was pooled and expressed as mean ± SEM and the P value determined by t test or ANOVA, *P < 0.01.