Figure 1: MAGE-A4 is a novel component of the RAD18 complex in cancer cells.

(a) Domain organization of full-length RAD18 and RAD18 Δ402–444 (which harbours an internal deletion removing the Polη-binding domain). (b) Spectral counts and estimated probability of true interaction by SAINT analysis for selected proteins identified in HA–RAD18-WT and Control (HA) APMS experiments. (c) Total protein signal intensity versus relative abundance between HA–RAD18-WT and HA–RAD18 Δ402–444 APMS. Signal intensity was normalized to the corresponding experiment’s bait intensity (x axis). (d) H1299 cells were infected with adenoviruses encoding WT HA–RAD18, HA–RAD18 Δ402–444 or with an ‘empty’ control adenovirus. Infected cells were treated with CPT (2 μM) or UVC (20 J m⁻²). Two hours (h) later, cell extracts were prepared and immunoprecipitated with anti-HA antibody-conjugated magnetic beads. The resulting immune complexes and input fractions were analysed by immunoblotting with anti-HA and anti-MAGE-A4 antibodies. (e) Expression vectors encoding MYC–RAD18, MYC–TRIM69 or green fluorescent protein (GFP) (for control plasmid) were transiently transfected into H1299 cells. Extracts from the resulting cells were immunoprecipitated with an anti-MYC antibody and the resulting immune complexes (or input fractions) were analysed by immunoblotting with antibodies against MAGE-A4 and MYC.