Journal: bioRxiv

Article Title: Interactome screening implicates BAG6 as a suppressor of UBQLN2 misfolding in ALS-dementia

doi: 10.1101/2025.10.15.682441

Figure Lengend Snippet: (A) co-IP of UBQLN2 and PEG10 from iPSCs expressing UBQLN2 WT , UBQLN2 P497H , or UBQLN2 4XALS with or without BAG6 knockdown. Immunoblotting with α-PEG10 antibodies revealed increased UBQLN2–PEG10 interaction in UBQLN2 P497H and UBQLN2 4XALS backgrounds, particularly under BAG6 knockdown conditions. (B) Quantification of PEG10 co-IP across genotypes demonstrates elevated PEG10 binding to UBQLN2 P497H and UBQLN2 4XALS compared to UBQLN2 WT . Statistical analysis of (A) was performed using two-way ANOVA with Tukey’s post hoc test. Data are presented as mean ± SEM from three independent experiments; *p ≤ 0.05, **p ≤ 0.01. (C) Reciprocal co-IP of UBQLN2 using α-PEG10 antibodies in UBQLN2 WT , UBQLN2 P497H , UBQLN2 4XALS , or UBQLN2 I498X iPSCs with or without BAG6 knockdown. (D) Quantification of UBQLN2 co-IP results from panel C demonstrated elevated PEG10 binding to UBQLN2 P497H and UBQLN2 4XALS compared to UBQLN2 WT under conditions of BAG6 knockdown. Statistical analysis of (C) was performed using two-way ANOVA with Tukey’s post hoc test. Data are presented as mean ± SEM from three independent experiments; **p ≤ 0.01, ***p ≤ 0.005. (E) Quantification of PEG10 RF1/2 levels from input lysates in (C), normalized to total PEG10 (RF1/2+RF1). Data were analyzed by two-way ANOVA with Tukey’s post hoc test. Values represent mean ± SEM from three independent experiments; ****p ≤ 0.0001.

Article Snippet: The following secondary antibodies were used for immunoblotting: IRDye 680RD goat anti-mouse (926-68070) and IRDye 800CW goat anti-rabbit (926-32211), both from LI-COR.

Techniques: Co-Immunoprecipitation Assay, Expressing, Knockdown, Western Blot, Binding Assay