Asymmetric Bidirectional Transcription from the FSHD-Causing D4Z4 Array Modulates DUX4 Production
Figure 4
Mutational analysis of the non-coding region of D4Z4.
(A) The non-coding region of D4Z4 with restriction enzyme sites used to construct various deletions is shown (no Δ) above the deletion series. Promoter regions involved in the various deletions are emphasized using alternating thickness of the rectangle and the name of the deletion is labeled at the right. (B) C2C12 cells were transfected with each mutant eGFP←D4Z4→dsRED and assayed for dsRED and eGFP expression by flow cytometry. The fluorescence intensity is plotted for the expression construct containing the most informative deletion (ΔNspI-AccIII) as well as mock transfected cells (Control) and cells transfected with the non-deleted vector (No Δ). (C) Quantification of the percentage of the total number of fluorescing cells that are dsRED-eGFP(+) for each mutation. Error bars = SD of the mean in three experiments that measured triplicate samples. * = p<0.05.