Fig. 7: S. suis facilitates DCAF15 deacetylation via SIRT1 to enhance EphA2 ubiquitination for disruption of blood-brain barrier. | Communications Biology

Fig. 7: S. suis facilitates DCAF15 deacetylation via SIRT1 to enhance EphA2 ubiquitination for disruption of blood-brain barrier.

From: Dcaf15-mediated EphA2 degradation triggers disruption of the blood-brain barrier during Streptococcus suis meningitis

Fig. 7: S. suis facilitates DCAF15 deacetylation via SIRT1 to enhance EphA2 ubiquitination for disruption of blood-brain barrier.The alternative text for this image may have been generated using AI.

A Co-IP analysis of acetylation of DCAF15 in hCMEC/D3 cells infected with WT SC19, Δstk or CΔstk. B Co-IP analysis of acetylation of exogenous DCAF15 in HEK293T cells co-transfected with Flag-DCAF15, HA-P300 in the treatment of deacetylase inhibitors TSA or NAM. C Co-IP analysis of ubiquitination of exogenous EphA2 in HEK293T cells co-transfected with Myc-EphA2, HA-Ub, Flag-DCAF15 in the absence or presence of NAM. D Co-IP analysis of acetylation of exogenous DCAF15 in HEK293T cells co-transfected with Flag-DCAF15 or HA-SIRT1, HA-SIRT2, HA-SIRT3, HA-SIRT4, HA-SIRT5 in the presence of NAM. E Deacetylation of DCAF15 in vitro from HEK293T cells co-transfected with HA-P300 and Flag-DCAF15 followed by  immunoprecipitated with Flag-agarose beads and then incubated with His-SIRT1. F Co-IP analysis of ubiquitination of EphA2 in HEK293T cells co-transfected with Flag-DCAF15, Myc-EphA2, HA-Ub under the condition of SIRT1 knockdown. G Western blot analysis of ZO-1 expression in SC19-infected hCMEC/D3 cells with SIRT1 knockdown. H Sequence alignment of the conserved K552 and K581 containing region in DCAF15 orthologs of different species. I, J Co-IP analysis of acetylation of DCAF15 or its mutants in HEK293T cells transfected with Flag-DCAF15 or its mutants including K552R, K560R, K581R (I) and DCAF15-2KR or DCAF15-2KQ (J) in the presence of NAM. K Co-IP analysis of the interaction between EphA2 and DCAF15 or its mutant (DCAF15-2KQ) in HEK293T cells transfected with Myc-EphA2, Flag-DCAF15 or its mutant. L Co-IP analysis of ubiquitination of EphA2 in SC19-infected hCMEC/D3 cells transfected with Flag-EV, Flag-DCAF15 or its mutants including DCAF15-2KR or DCAF15-2KQ. M ZO-1 expression in SC19-infected or non-infected hCMEC/D3 cells transfected with Flag-DCAF15 or its indicated mutants. N The TEER value in transwell infection model transfected with Flag-EV, Flag-DCAF15 or its indicated mutants. Data are representative of three independent experiments with triplicate samples per group. All data are presented as mean ± SD. P ≤ 0.05 was considered as statistical significance.

Back to article page