Fig. 2

4q35 gene expression is affected by epigenetic drugs depending on 4q allele size. Expression data of 4q35 genes in human primary myoblasts (HPMs) (A–F) and in human trophoblasts cells (HTCs) (G-L) carry a normal sized allele (CTRL (> 10U)) or D4Z4 reduced alleles (DRA:7U and DRA:4U). Cells were treated or not treated with the indicated compounds: 5-Aza-2′-deoxycytidine (5-Aza-dC), trichostatin A (TSA), nicotinamide (NAM), PARP inhibitor (PJ34). SLC25A4 (A,G), FAT1 (B,H), FRG1 (C,I), FRG2 (D,J), DBE-T (E,K) and D4Z4-T (F,L) RNAs were measured by RT-qPCR and normalized over the RPLP0 reference gene. The values of the CTRL samples (n = 3) and FSHD DRA:4U (n = 2) were averaged. Experiments were done in triplicate, and the results were analyzed using two-way ANOVA tests to perform multiple comparisons. Hashtags (#) indicate the statistical significance of data from treated samples compared to untreated samples (NT) in each group. Asterisks (*) indicate statistical significance of data from treated cells carrying DRA compared to the same treatment in control cells. Plus signs ( +) indicate statistical significance of data from DRA:7U compared to DRA:4U cells. P value ranges are as follows: *, #, + 0.05 < p-value < 0.01; **, ##, + + 0.01 < p-value < 0.001; ***, ###, + + + 0.001 < p-value < 0.0001; ****, ####, + + + + p-value < 0.0001