Fig. 2
From: Familial STAG2 germline mutation defines a new human cohesinopathy
Sister chromatid cohesion assay and immunoreaction experiments. a p.Ser327Asn STAG2 does not show effect on the sister chromatid phenotype. Open (gray) and closed (black) sister chromatids are shown for the patient and four controls with WT STAG2 (C1-4). A total of 100 metaphases were analyzed for each individual. The amount of closed sister chromatids are indicated at the bottom of each individual column. Open and closed sister chromatid phenotypes in metaphase cells from the patient are demonstrated. b Immunofluorescence of patient fibroblasts. Cultured fibroblasts from the proband were incubated overnight with anti-STAG2 (J-12) at 4 °C followed by Alexa 488-conjugated second antibody (green) and nuclear counterstain with DAPI (blue). The nuclear localization of STAG2 was confirmed in the double label immunofluorescence merged picture. Similar results were observed in three independent experiments. c Western blot of STAG2 from patient fibroblasts. Protein extracts from patient and control individuals were subjected to immunoblotting using the anti-STAG2 (J-12). After visualization of STAG2, the blot membrane was stripped and re-probed with alpha-tubulin antibody, which was used as a quantitative control. Similar results were observed between the patient and the two controls in three independent experiments
