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. 2019 Nov;157(5):1398–1412.e9. doi: 10.1053/j.gastro.2019.07.029

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© 2019 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Mecp2 phosphorylation at S80 is required for HSC proliferation and collagen expression. (A) Western blot for Mecp2, pMecp2^S80, pMecp2^S421, HIPK2, α-SMA, and GAPDH in HSCs after 0, 2, 5, and 8 days in culture. (B) Proliferation assay: BrdU incorporation was measured in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. (C) Viability assay: reduced MTT was measured in Mecp2^S80A HSCs (compared with WT HSCs) after 0, 2, 5, and 8 days in culture. (D) mRNA level of Mcm5 quantified by qRT-PCR in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. Western blot for MCM5 and β-actin in WT and Mecp2^S80A HSCs after 8 days in culture. (E) mRNA level of CCNA2 and BRIP1 in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. (F) Western blot cyclin D1 and E1 in WT and Mecp2^S80A activated HSCs (8 days in culture). (G) Col1A1, α-SMA, and TIMP1 quantified by qRT-PCR in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. Error bars represent mean ± SEM. Statistical significance was determined by ANOVA or Students t test; *P < .05, ** P < .01, *** P < .001. denotes a trend with P < .1.

Inline graphic — Mecp2 phosphorylation at S80 is required for HSC proliferation and collagen expression. (A) Western blot for Mecp2, pMecp2^S80, pMecp2^S421, HIPK2, α-SMA, and GAPDH in HSCs after 0, 2, 5, and 8 days in culture. (B) Proliferation assay: BrdU incorporation was measured in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. (C) Viability assay: reduced MTT was measured in Mecp2^S80A HSCs (compared with WT HSCs) after 0, 2, 5, and 8 days in culture. (D) mRNA level of Mcm5 quantified by qRT-PCR in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. Western blot for MCM5 and β-actin in WT and Mecp2^S80A HSCs after 8 days in culture. (E) mRNA level of CCNA2 and BRIP1 in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. (F) Western blot cyclin D1 and E1 in WT and Mecp2^S80A activated HSCs (8 days in culture). (G) Col1A1, α-SMA, and TIMP1 quantified by qRT-PCR in WT and Mecp2^S80A HSCs after 0, 2, 5, and 8 days in culture. Error bars represent mean ± SEM. Statistical significance was determined by ANOVA or Students t test; *P < .05, ** P < .01, *** P < .001. denotes a trend with P < .1.