Fig. 4: Impact of CDC42 variants on ER stress. | Nature Communications

Fig. 4: Impact of CDC42 variants on ER stress.

From: Autoinflammatory patients with Golgi-trapped CDC42 exhibit intracellular trafficking defects leading to STING hyperactivation and ER stress

Fig. 4

A Immunofluorescence analyses of ER stress in fibroblasts from the HD16 healthy donor and from the Y64C and R186C CDC42 patients. Left: co-stainings for BiP, F-actin and nuclei are shown. Images are representative of 3 independent experiments. Scale bars: 10 µm. Right: quantification of microscopy images. One dot represents the mean value from about 15 cells from one independent experiment. Results are shown as means +/- SEM of three biological replicates and the significance levels were calculated using ordinary one-way ANOVA (*: P = 0.0358). B Western blot analysis of BiP and CDC42 expression in HD8, Y64C and R186C fibroblasts treated with DMSO (vehicle, -) or thapsigargin (+). GAPDH is shown as a loading control. The Molecular Weights (kDa) are indicated. C, Flow cytometry analyses of BiP expression in THP-1 cells expressing WT or mutant CDC42. Left: examples of BiP expression FACS profiles in cells treated with DMSO (vehicle; turquoise) or thapsigargin (red). Right: quantifications of the BiP Mean Fluorescence Intensity (MFI) in cells treated with DMSO (-) or thapsigargin (+). One dot represents the MFI from one independent experiment. Results are shown as means +/-  SEM from three biological replicates and the significance levels were calculated using two-way ANOVA (*P < 0.0359). D Expression levels of HSPA5 (left) and ATF4 (right) mRNA in healthy donors (HDs) or CDC42 patients’ fibroblasts treated with DMSO (-) or thapsigargin (+). E Expression levels of HSPA5 (left), ATF4 (center), and DDIT3 (right) mRNA in THP-1 cells expressing WT or variants CDC42 upon DMSO (-) or thapsigargin (+) treatment. NI: non infected. For panels D and E, each dot represents the mean value of three technical replicates from one independent experiment. Results are shown as means +/- SEM from at least three biological replicates and the significance levels were calculated using ordinary one-way ANOVA (*P < 0.0328; **P < 0.0081; ***P < 0.0004; ****P < 0.0001). Source data are provided as a Source Data file.

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