Fig. 2
ERCC4/XPF/FANCQ (missense) mutations and their effects. a: Dose-response curves of FA-Q fibroblasts 3104 (red) compared with previously reported 1333 (blue), XFE (XP51RO; orange) and XP-F fibroblasts (XP23OS; green) reveal different degrees of UV-C sensitivity on survival compared with normal control fibroblasts (black) (means ± SEMs). LC50 levels are indicated by the dashed lines of corresponding colors; they equal 5.7 J/m2 for control cells, 2.8 J/m2 for 3104, 1.5 J/m2 for 1333, 2.2 J/m2 for XP23OS and 1.3 J/m2 for XP51RO. All mentioned cells represent primary fibroblasts. b: NER activity of 3104 FA-Q primary fibroblasts assayed ex vivo. UDS (upper panel) was measured by incorporation of 5-ethynyl-deoxyuridine (EdU), RRS (lower panel) by incorporation of 5-ethynyl-uridine (EU). Mixed-in wildtype (WT) fibroblasts preloaded with polystyrene beads (not depicted) served as internal control. DAPI as nuclear counterstain for all cells. UDS and RRS signals were quantified from 20 to 40 random nuclei. c: UDS levels of control fibroblasts (grey) compared to FA-Q 3104 (red), 1333 (blue), XFE (orange) and XP-F (green) fibroblasts (means ± SEMs, brackets indicates levels os significance). d: RRS levels of control fibroblasts (grey) compared to FA-Q 3104 (red), 1333 (blue), XFE (orange) and XP-F (green) fibroblasts (means ± SEMs, brackets indicates levels os significance). e: Dose-response curves of 3104 (red; means ± SEMs) and two previously reported FA-Q lymphoblast lines (FA104, green; 1333, blue) exposed to MMC show different degrees of sensitivity compared to normal control lymphoblasts (black). LC50 levels are indicated by dashed lines of corresponding colors and equal 210 nM MMC for the control, 80 nM MMC for 1333, 42 nM MMC for 3104 and 24 nM MMC for FA104. f: Combinations (brackets) of the biallelic mutations in all XP-F, CS, XPCS(FA) and XFE patients with p.Arg589Trp (shaded) (above) and of the biallelic mutations in the three reported FA-Q patients (below) are depicted relative to location in the ERCC4/XPF/FANCQ protein domain structure [14]. The predicted mutational effects on the protein level are shown in bold letters together with the corresponding patient designations listed below. Individuals with XP-F include XP24BR, XP32BRand AS871, with XFE include XP51RO, with XPCS(FA) include XPCS1CD, and with FA-Q include 3104, FA104 and 1333 (3104 highlighted in red). The phenotypic severity of disease in patients with XP, CS, XPCS(FA), or FA-Q carrying one heterozygous p.Arg589Trp mutation is indicated above the brackets