Fig. 3

Smarcad1 reinforces Trim28 binding and allows H3.3 accumulation and heterochromatinization of proviral sequences. A ChIP‒qPCR was performed using Trim28 antibody on day 2 (n = 3) and 7 (n = 6) after MLV-PBSpro infection, with primers for MLV provirus-specific regions and (B) for open and repressed genomic chromatin loci, including ERVs (n = 12). Values shown are mean ± s.e.m, relative to the total input samples and normalized to the signal of negative control gene (Gapdh). A control with IgG antibody gave background enrichment. C ChIP‒qPCR was performed using HA antibody, on day 2 (n = 3) and 7 (n = 5) after MLV-PBSpro infection and 8 h Dox induction, with primers for MLV provirus-specific regions and D for open and repressed genomic chromatin loci, including ERVs (n = 12). Values shown are mean ± s.e.m, relative to the total input samples and normalized to the signal of the negative control gene (Hbb). A control without HA induced expression (no Dox) gave background enrichment. In all panels, Mann–Whitney U Test was used for statistical analysis of difference from IgG background enrichment (red asterisks) or between the shControl and shSmarcad1 samples (black); *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001