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. 2020 Aug 31;130(10):5272–5286. doi: 10.1172/JCI134966

Figure 2. Expression and localization of MCM10 mutants.

Figure 2

(A) Primary fibroblasts from proband or healthy donor were lysed and probed for MCM10 (left). Intensity of MCM10 was normalized to loading control (actin, right). (B) Western blot of stable SFB-MCM10 expression in 293T whole cell extracts. Full-length SFB-MCM10 and truncation mutants were detected with anti-FLAG antibody (top) and with anti–α-tubulin antibody as a loading control (bottom). (C) Confocal imaging of SFB-MCM10 localization in stable 293T cell lines. Full-length SFB-MCM10 and truncation mutants were detected with anti-FLAG antibody (green) with DAPI staining (blue). Scale bars: 10 μm. (D) WT GFP-MCM10 or GFP-R582X MCM10 were transiently expressed in 293T cells and imaged by confocal microscopy with quantification of nuclear GFP (right). Mean ± 95% CI. ***P < 0.001, Kruskal-Wallis with Dunn’s multiple comparison test. n = 48 (WT); n = 55 (R426C); n = 40 (R582X). Scale bars: 10 μm. Data representative of 3 technical replicates.