Fig. 3: C2C12 cells overexpressing Tet1s show aberrant 5mC oxidation and higher levels of LINE 1 ORF1p.

A Cells transfected with EGFP-Tet1, EGFP-Tet1s or EGFP alone, were immunostained against 5hmC. Fluorescence intensity levels of overexpressed proteins and 5hmC were measured, sum nuclear 5hmC levels were normalized to the sum nuclear DAPI intensity and grouped as described in Supplementary Fig. 2B, D. B C2C12 cells non-transfected and transfected with GFP-Tet1s were synchronized and immunostained against LINE 1 ORF1 protein. Fluorescence intensity levels of the protein were measured and mean cytoplasmic levels were plotted. Representative confocal images for LINE 1 ORF1p and TET1/TET1s immunostainings are shown in C. For all boxplots, the box represents 50% of the data, starting in the first quartile (25%) and ending in the third (75%). The line inside represents the median. The whiskers represent the upper and lower quartile. Statistical significance was tested with a paired two-samples Wilcoxon test (n.s. not significant, is given for p-values ≥ 0.05; one star (*) for p-values < 0.05 and ≥ 0.005; two stars (**) is given for values < 0.005 and ≥ 0.0005; three stars (***) is given for values < 0.0005). N-numbers and p-values are shown in Supplementary Data 1. Source data are provided as a Source Data file. Scale bar = 5 μm.