Fig. 1: TDP-43 interacts in vivo and in vitro via RRM1 and RRM2 domains, with HDAC1 that can module its acetylation.

a Spinal cord, cerebellum, or striatum of BALB31c mice were used for co-immunoprecipitation experiments. After dissection tissues were lysed and protein extracts were immunoprecipitated, with specific antibodies. Proteins retained were separated on SDS-PAGE and visualized by western blot using specific antibodies (b, c) HEK 293T cells were transiently transfected with FLAG-tagged HDCA1 and Myc-tagged WT or mutant TDP-43. We tested pathological mutants M337V, A382T, D169G, K263E, acetylation null KK-AA or mimicking KK-QQ mutants, and the deletion mutants ΔRRM1, ΔRRM2, ΔG-rich, ΔRRM1/RRM2. Forty-eight hours after transfection, cells were lysed, protein extracts were immunoprecipitated and analysed as in (a). d Bar graph shows the relative binding of HDAC1 to mutant TDP-43, normalized to TDP-43 WT. The data were obtained from four independent experiments; *p > 0.05 and **p > 0.01 versus WT binding, analysed by using one-way ANOVA with Bonferroni’s Multiple comparison post-hoc test. e Representative 2-DE maps showing the change in TDP-43 acetylation status, upon HDAC1 or HDAC6 expression. SH-SY5Y were transfected by TDP-43 WT or deletion mutant ΔRRM1-2 and FLAG-tagged HDCA1 or HDAC6. Forty-eight hours after transduction, TDP-43 was immunoprecipitated and a visualized by western blot using an anti-acetyl lysine antibody.