Figure 2.

Adenine base editing of the DUX4 PAS reduces expression of DUX4 and its target genes in FSHD myogenic cells derived from polyclonal cultures

(A) mRNA levels as assessed by qRT-PCR of DUX4 and four DUX4 target genes (MBD3L2, ZSCAN4, TRIM43, and KHDC1L) in PAS unedited versus edited clones derived from two FSHD1 and one FSHD2 cell lines differentiated into myotubes. Statistical significance was calculated with unpaired two-tailed t test (ns, non-significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001) on log2 transformed expression values to correct for skewed distribution. Expression values normalized to GUSB as housekeeping gene are plotted. Line represents mean, and whiskers represent min and max value. Individual dots represent individual clones; the two violet clones carry a deletion affecting the DUX4 PAS. (B) mRNA levels of two myogenic markers (MYOG and MYH3) for all unedited and edited clones of all three FSHD cell lines are plotted. Statistical significance was calculated with unpaired two-tailed t test (ns, non-significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). Bars represent mean ± SEM, with individual clone expression values plotted as individual dots.