Fig. 1

SIRT5 binds to and desuccinylates ANXA1 at K166 after OGD/R. A Primary cultured microglia were subjected to oxygen and glucose deprivation/reoxygenation (OGD/R) for 24 h. Coimmunoprecipitation (Co-IP) analysis indicating the succinylation level of ANXA1. B HEK293T cells were transfected with plasmids encoding Flag-tagged SIRT5 and SIRT7 together with haemagglutinin (HA)-tagged ANXA1. Co-IP analysis showing the succinylation level of ANXA1. C HEK293T cells were transfected with HA-ANXA1 and subjected to SIRT5 knockdown. Co-IP analysis showing the succinylation level of ANXA1. D Co-IP analysis showing the succinylation level of ANXA1 in primary cultured microglial cells subjected to SIRT5 knockdown. E HEK293T cells were transfected with plasmids encoding HA-ANXA1 and Flag-tagged wild-type SIRT5 (SIRT5-WT) or SIRT-H158Y (SIRT5-HY). Co-IP analysis was conducted to show the succinylation level of ANXA1. F HEK293T cells were transfected with plasmids encoding HA-ANXA1-WT, K97R, K161R, K166R, K195R, or K312R mutants, respectively. The cell extracts were collected for IP with anti-HA beads, followed by IB analysis with the indicated antibodies and showing the succinylation level of ANXA1. Data represent at least three independent experiments. M_r, relative molecular mass