Fig. 9: In cellula assessment of TMX4 engagement in mixed disulfides with NESPRIN3α. | Nature Communications

Fig. 9: In cellula assessment of TMX4 engagement in mixed disulfides with NESPRIN3α.

From: TMX4-driven LINC complex disassembly and asymmetric autophagy of the nuclear envelope upon acute ER stress

Fig. 9: In cellula assessment of TMX4 engagement in mixed disulfides with NESPRIN3α.The alternative text for this image may have been generated using AI.

a Lysates of cells mock-transfected (lanes 1, 5, 9), expressing GFP-SUN1 and HALO-NESPRIN3α, TMX4C67A-V5 (lanes 2, 6, 10), GFP-SUN1 and HALO-NESPRIN3α (lanes 3, 7, 11), or TMX4-V5 GFP-SUN1 and HALO-NESPRIN3α (lanes 4, 8, 12) were separated in a reducing gel and transferred on PVDF membranes. Expression of the ectopic proteins was confirmed by WB with anti-V5 (lanes 1–4), anti-GFP (lanes 5–8), or anti-HALO antibodies (lanes 9–12). Uncropped blots in Supplementary Fig. 6. b The presence of TMX proteins (lanes 1–3), NESPRIN3α (lanes 4–6), or SUN1 (lanes 7–9) in mixed disulfides (MD) with TMX4C67A-V5 (lanes 2, 5, 8) or TMX4-V5 (lanes 3, 6, 9) immunoisolated from cell lysates with anti-V5 antibodies was monitored under non-reducing conditions by western blot with anti-V5, anti-HALO, or anti-GFP antibodies, respectively. c. The engagement of TMX proteins (lanes 10–12), or NESPRIN3α (lanes 13–15) in MD with HALO-NESPRIN3α immunoisolated from cell lysates with anti-HALO antibodies was monitored under non-reducing conditions by western blot with anti-V5 and anti-HALO antibodies, respectively. d The presence of TMX4 proteins in MD with GFP-SUN1 (lanes 1–3), or with HALO-NESPRIN3α (lanes 4–6) immunoisolated from cell lysates has been assessed by WB with anti-V5 antibodies under non-reducing conditions (or reducing conditions, lanes 7–12). WB is representative of at least three independent experiments.

Back to article page