LMNA(R482W) elicits H3K27 acetylation of predicted MIR335 enhancers after induction of differentiation. (A and B) H3K27me3 (A) and H3K27ac (B) enrichment determined by ChIP-qPCR upstream of MIR335 in undifferentiated and differentiated (day 3) ASCs, ASC^LMNAWT, and ASC^LMNA(R482W) (means ± SD of duplicate transductions and differentiations, each analyzed by duplicate ChIP). Values for IgG ChIPs for all regions and cell types ranged from 0.02 to 0.24 and are not depicted. Statistics for A: one-way ANOVA on r1–r4 Undiff., P = 0.306; r6–r11 Undiff., P = 0.169; r1–r4 Diff., P = 0.200; and r6–r11 Diff., P = 0.001. ^§P < 0.01, Fisher’s exact tests relative to the same undifferentiated cell type: *, P < 0.05; **, P < 0.01; post-ANOVA t tests. (B) One-way ANOVA on r1–r4 Undiff., P = 0.300; r6–r11 Undiff., P = 0.001; r1–r4 Diff., P = 0.002; and r6–r11 Diff., P < 0.001; ^§P < 0.01, Fisher’s exact tests relative to the same differentiated cell type; *, P < 0.05; **, P < 0.01; post-ANOVA t tests. (C) Dual-color FISH analysis of the MIR335 gene (red signals) and MIR335 enhancer regions r10 and r11 (green signals) in native ASCs, ASC^LMNAWT, and ASC^LMNA(R482W) before and after a 3-d adipogenic induction. Representative views are shown. Bars: (main images) 5 µm; (insets) 0.5 µm. (D) Quantification of FISH observations (n = 44–64 alleles analyzed in two independent FISH analyses; means ± SD; **, P = 0.0082; Fisher’s exact test).