Fig 1: Metabolic genes are altered in Mecp2/Y AE2 cells due to loss of MECP2- and NCOR1/2-mediated transcriptional repression. (A) t-SNE plot of +/Y and Mecp2/Y AE2 cells after single-cell RNA sequencing, n = 2000 cells, collected at P18. (B) Volcano plot showing genes with altered expression in Mecp2/Y AE2 cells after single-cell RNA sequencing. The top 20 significantly altered genes are labeled. Significant genes have −log10(P-value) < 0.05. (C) SREBP1 expression in the +/Y and Mecp2/Y lung at P21 and P56. The 68 kDa band (shown) is the active, cleaved protein, whereas the 120 kDa inactive precursor was not visualized. GAPDH was used for normalization. (D) TBL1XR1 is associated with NCOR1/2 co-repressor complex members NCOR1, MECP2 and HDAC3 in the mouse lung, as shown by co-immunoprecipitation (co-IP) assay. This complex forms without MECP2 in Mecp2/Y lung tissue. i, input (nuclear protein); ip, immunoprecipitated protein. Anti-TBL1XR1 and anti-H3K4me3 (positive control) ChIP-qPCR of sites surrounding the (E) Acot1 promoter and (F) Hmgcs1 promoter, n = 3. Data are expressed as mean ± SEM. Statistics were assessed using Student’s t-test. *P < 0.05, **P < 0.01. FC, fold change.