Fig. 5: WFS1 is a transmembrane protein of the ER partially located in the inner nuclear membrane.
From: MyoD1 localization at the nuclear periphery is mediated by association of WFS1 with active enhancers
a Schematic drawing of the predicted structure and orientation of WFS1 transmembrane protein. Positions of specific tags in ectopically expressed constructs are indicated. NH2: N-terminus, COOH: C-terminus, ER: Endoplasmic Reticulum (b) Immunofluorescence analysis of WFS1 cellular localization in indicated genotypes. Scale bars, 10 µm. c WFS1 Western blots of total cell extracts (input) and of immunoprecipitated WFS1 (IP samples) using WFS1 antibody or IgG as control. Orange star marks unspecific band. d Representative confocal immunofluorescence images of WFS1 knockout cells ectopically expressing WFS1 tagged with FLAG, HA and mScarlet as indicated in (a), following membrane permeabilization with Triton X-100 (all membranes) or digitonin (plasma membrane only), using antibodies to the indicated antigens (anti-LAP2α and anti-HA tag). mScarlet was either visualized by its own fluorescence (second panel) or by using an antibody (Ab) recognizing mScarlet (first panel). Scale bars, 10 µm. e WFS1 knockout cell clone ectopically expressing FLAG-tagged WFS1 was processed for super-resolution immunofluorescence microscopy using antibodies against WFS1 and LAP2β or lamin A/C. Representative super-resolution images for LAP2β- and WFS1- co-stained cells (AiryScan) are shown. Normalized signal intensity of WFS1 and LAP2β or lamin A/C was determined using line profiles across co-stained nuclei. Graphs display the relative localization of WFS1 to INM protein LAP2β (top) and intranuclear protein lamin A/C (bottom). Circles mark magnified areas of the graphs displayed on the left. Displayed are mean values ± SEM (ntop = 25, nbottom = 25). Scale bars, 5 µm. Images processed in Fiji. Source data are provided as a Source Data file.
