Fig 1: Distinct MUNC spliced domains are required for binding to specific genomic sites, regulating adjoining gene expression, and interacting with protein partners(A) Heatmap of ChIRP-qPCR (“MUNC binding”) and RT-qPCR (“gene expression”) Z scores upon expression of MS constructs in proliferating cells. Mean values from three biological replicates are presented as Z score. Green outlined boxes identify domains required both for MUNC binding and gene expression regulation.(B and C) Left: scheme of point mutations in CH1 (B) and CH4 (C) domains (with compensatory mutations to maintain base-pairing when necessary). Right: RT-qPCR of Myod1, Myog, and Myh3 mRNAs normalized to Gapdh upon expression of MUNC constructs in proliferating cells. Shown relative to MS. Three independent transfectants (mean ± SEM).(D) SMC3 association with MUNC constructs in proliferating cells. Levels of MUNC lncRNA in RIP-qPCR were normalized to H19 lncRNA and shown relative to MS. Three biological replicates (mean ± SEM). *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed Student’s t test.See also Figures S6 and S7.
Fig 2: Distinct structural domains of the spliced isoform of MUNC are required for induction of promyogenic factors.(A) Constructs used in the study.(B–D) RT-qPCR analyses of (B) Myod1, (C) Myog, and (D) Myh3 mRNAs normalized to Gapdh and shown relative to MS overexpression in proliferating control cells. Data are representative of at least two independent transfectants with at least three biological replicates for each of them (mean ± SEM).(E–G) Plots of average induction of (E) Myod1 versus Myog, (F). Myod1 versus Myh3, and (G) Myog versusw Myh3 in Ctrl cells and cells expressing MS constructs. Black line is the theoretical correlation line if the expression of one gene is dependent on another; blue line shows experimental correlation.(H) RT-qPCR analysis of MUNC levels normalized to Gapdh and shown relative to Ctrl. Three biological replicates (mean ± SEM).(I–K) RT-qPCR analyses of (I) Myod1, (J) Myog, and (K) Myh3 mRNAs normalized to Gapdh and shown relative to MS. Data are representative of three biological replicates (mean ± SEM). *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed Student’s t test.See also Figure S4.
Supplier Page from Proteintech Group Inc for MYH3 antibody