(A) Top: Lysates of HK2 cells overexpressing RTN1A or RTN1C were immunoprecipitated (IP) with anti-HK1 antibody or control IgG and immunoblotted with VDAC1 antibody. Bottom: Input lysates were immunoblotted with VDAC1, RTN1A, and RTN1C antibodies. (B) Immunoprecipitation was performed using purified recombinant proteins (DDK-tagged HK1, His-tagged VDAC1, and Myc-tagged RTN1A). Interaction of HK1 and VDAC1 was assessed with increasing amounts of RTN1A (B) or RTN1C (C) using anti-His antibody and immunoblotted with DDK and Myc antibodies. (C-D) Western blot and densitometric analysis of Caspase-1 in HK2 cells with RTN1A or RTN1C overexpression. (D-G) Western blot and densitometric analysis of Caspase-1 and IL-1β HK2 cells overexpressing RTN1C or RTN1A with or without HK1 overexpression. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 between indicated groups by one-way ANOVA with Tukey's post hoc analysis.