Fig 3
Matrigel and low confluency conditions produced the highest editing efficiency. A Compiled results for all editing experiments. Replicates from three skMDC lines were transfected with one of three guides under different conditions. B Correlation between editing efficiency and the number of cells at the time of transfection. There is a clear negative relationship between cell count and editing efficiency under Matpos conditions (R2 = 0.75; p = 0.0012). n per condition = 5 (1× 18M, 1× 32F, 3x 42F). C Comparison of editing efficiency between guides within each condition. Different skMDC donor cell lines are indicated with different symbols. Experiments using Lo/Matpos conditions were repeated to confirm our findings. D Correlation between the percentage of ATTO 550+ cells and MYOD1 editing efficiency in three skMDC lines transfected under different conditions. E Correlation between MFI and MYOD1 editing efficiency in three skMDC lines transfected under different conditions. All data points represent individual replicates. Error bars show mean ± SD. ** = p < 0.01; *** = p < 0.001; **** = p < 0.0001. NC = negative control; Lo = low confluency, Hi = high confluency; Matneg = uncoated wells; Matpos = Matrigel coated wells; MFI = median fluorescence intensity