Figure 2

DJ-1 inhibited MAP1b-LC aggregation in HEK293t and SH-SY5Y cells. A, Flag-MAP1b-LC was co-transfected with HA-DJ-1 or pcDNA3 vector into HEK293t cells for 48 hrs before the cells were lysed and separated into Triton x-100 soluble and insoluble fractions. Western blot results showed overexpression of DJ-1 almost abolished the insoluble Flag-MAP1b-LC. B, Flag-MAP1b-LC and HA-DJ-1 or pcDNA3 were co-transfected into HEK293t cells for 48 hrs before the cells were fixed and immuno-stained with rabbit anti-Flag or mouse anti-HA antibodies. The most representative images were shown and the results showed the inhibition of MAP1b-LC aggregates in the HA-DJ-1 transfected cells. C, Flag-MAP1b-LC was co-transfected with pcDNA3-HA-DJ-1or pcDNA3 into the SH-SY5Y cells and cultured for 48 hrs before the cells were lysed and separated into Triton X-100 soluble and insoluble fractions. Western blot results showed that overexpression of DJ-1 suppressed the insoluble Flag-MAP1b-LC. D, SH-SY5Y cells were transfected with Flag-MAP1b-LC and pcDNA3-HA-DJ-1 or pcDNA3 respectively. Forty-eight hours later the cells were fixed and stained with rabbit anti-Flag or mouse anti-HA. The results showed the inhibition of MAP1b-LC formed aggregates in the HA-DJ-1 transfected cells. E, pcDNA3-HA-DJ-1 or pcDNA3 was transfected into the SH-SY5Y cells for 48 hrs before the cells were lysed and separated into Triton x-100 soluble and insoluble fractions. Western blot results suggested overexpression of DJ-1 could suppress the endogenous insoluble MAP1b-LC.