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. Author manuscript; available in PMC: 2011 Feb 1.
Published in final edited form as: Gastroenterology. 2009 Oct 17;138(2):705. doi: 10.1053/j.gastro.2009.10.002

Figure 4. MeCP2 expression is regulated by miR132.

Figure 4

(a) Micro RNAs were isolated from rat qHSC and myofibroblasts using miRNeasy mini kit and reverse transcribed using miScript Reverse Transcription Kit. miR132 was detected with miScript primer assay 218300. (b) Micro RNA was purified from HSC isolated from livers of mice following BDL (or sham control) and CCl4 (or olive control) injuries and miR132 detected as described above. (c to e) 5×106 myofibroblasts were mixed with 2μg miR132 mimic or control siRNA and electroporated. Cells were harvested 48 hours later for RNA and whole cell protein extract. (c) mIR132 was quantified by miScript primer assay 218300 (d) Whole cell protein extracts were separated by SDS-PAGE and immunoblotted for MeCP2 and β–actin. (e) cDNA obtained in (b) was further used as template for qRT-PCR analysis of PPARγ. (f) Native chromatin was prepared from in vitro transdifferentiated Wt or Mecp2-/y myofibroblasts. 100μg of native chromatin was incubated with 10μg of anti-dimethyl H3K27, H3K9 or H3K4 and ChIP assays carried out. Immunoprecipitated DNA was used as template in qPCR reactions using mouse PPARγ promoter specific primers.