Journal: iScience
Article Title: Loss of DDI2 rewires proteostasis through CCN1-driven compensatory autophagy
doi: 10.1016/j.isci.2026.115056
Figure Lengend Snippet: DDI2 interacts with CCN1 and p97 (A) MRC5 and MIA PaCa-2 cells, either DDI2 KO or control, are treated with DMSO or CB-5083 (10 μM for 3 h). CCN1 protein levels are analyzed by immunoblotting using antibodies specific to DDI2, CCN1, and β-Actin, and representative blots with corresponding densitometric bar graphs are shown. (B) HEK293 cells transiently expressing protease-dead DDI2 (Flag-dRVP DDI2) are subjected to immunoprecipitation using anti-FLAG beads, followed by immunoblotting with antibodies specific to DDI2, CCN1, p97, and RAD23A. Lysate lanes are loaded with 5% of the total input used for immunoprecipitation. The molecular weights of the proteins analyzed are as follows: P97/VCP (89 kDa), RAD23A (52 kDa). (C) Representative confocal immunofluorescence microscopy images show the co-localization of DDI2 (red) and CCN1 (green) in wild-type MRC5 and MIA PaCa-2 cells. The nuclei are visualized by staining with DAPI (blue). Scale bars, 10 μm. Three biological replicates for each cell line are used to perform Western blotting. Statistical significance of each condition compared to the indicated control or treatment is determined using unpaired Student’s t test or two-way ANOVA with Šídák’s post hoc test, as appropriate. Data are represented as mean ± SEM. Significance levels are indicated as follows: ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001. Corresponding immunoblots, including molecular weight marker lanes, are provided in .
Article Snippet: CB-5083 , Selleckchem , Cat#S8101.
Techniques: Control, Western Blot, Expressing, Immunoprecipitation, Immunofluorescence, Microscopy, Staining, Molecular Weight, Marker
