Fig. 3

Reduced DAP1 protein and enhanced autophagy in DAP1 SLE risk genotype. B cell-derived lymphoblastoid cell lines (LCLs) on 8 participants from 1000 genome study were purchased from Coriell based on their DAP1 genotypes. a, b Lymphoblastoid cells (LCLs) from four donors with protective genotype (TT) or four donors with risk genotype (CC) incubated with RPMI in the absence (a) or presence of 100 nM bafilomycin A1 (b) for 4 h. Cell lysates were analyzed by western blot using indicated antibodies. The ratio of DAP1/GAPDH (c) or the ratio of LC3-II/LC3-I (d) or the ratio of p62/GAPDH (e) is presented in bar graphs. Data are representative from at least three independent experiments. f Show quantitative RT-PCR analysis of DAP1 mRNA in LCLs from four donors with protective genotype TT or four donors with risk genotype CC. GAPDH was used as an internal control to normalized DAP1 expression. All plotted values are averages of two independent experiments using two different primer pairs. g Gating strategy and flow plots from CYTO-ID green autophagy detection assay on 4 TT or 4 CC LCL samples. Bar graph represents the average ± SEM of two independent experiments. Error bars: SEM; *p < 0.05. ns, not significant. Student’s t test