Fig 2: Effect of short-term diabetes on KIF5B. a RNA was extracted from DRG of control (Ctrl) and diabetic (streptozotocin, STZ) male (M) and female (F) rats. RT-PCR was used to analyze mRNA levels of KIF5B. b KIF5B protein levels analyzed by immunoblotting in DRG total extracts of Ctrl and STZ M and F rats. c KIF5B protein levels analyzed by immunoblotting in isolated axoplasm of Ctrl and STZ M and F rats. Representative Western blots are presented above the graphs, with the respective loading controls (GAPDH). The densitometry of each band was analyzed and the results are expressed as percentage of M Ctrl. The columns represent the mean ± SEM, n = 6 animals for each group. The effects of sex, diabetes, and the interaction sex by diabetes were analyzed using two-way ANOVA followed by Bonferroni post hoc test (significance: *p < 0.05; **p < 0.01)

Fig 3: Brucella infection dissociates BORC-related lysosome trafficking factor KIF1b but recruits KIF5b.Colocalization of KIF1b with BCVs or LAMP1 in the infected control and mBloc1s1 (A), or in wt-Bloc1s1 and Rr-Bloc1s1 (B) cells at the indicated h.p.i. Colocalization of KIF5b with BCVs or LAMP1 in the infected control and mBloc1s1 (C), or in wt-Bloc1s1 and Rr-Bloc1s1 (D) cells at the indicated h.p.i. White arrows: colocalization of BCVs with the indicated proteins. Insets: magnification of the selected areas (within windows with dash white lines). Yellow and orange arrows: the perinuclear and peripheral accumulation of BCVs-Lamp1, respectively. Bar: 5 µm. Quantification of BCV-KIF1b+ (E) and BCV-KIF5b+ (F) in Bm16M-infected cells at the indicated h.p.i. showing in A, B and C, D, respectively. Dynamics of LAMP1-KIF1b+ (G) or LAMP1-KIF5b+ (H) in a time course (48 hr) of Bm16M infection at the indicated h.p.i. Host cells were infected with or without Bm16M, and at the indicated h.p.i., the cells were fixed and subjected to confocal immunofluorescence assays. Images are representative of three independent experiments. Statistical data represent means ± standard error of mean (SEM) from three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001.

Fig 4: TLR7-mediated autophagy modulation of the expression of KIF5A.a–d Representative images of Kinesin family member 5 family member (KIF5A, KIF5B, and KIF5C) expression in the hippocampus were detected by western blot (n = 6). Quantitative analysis was performed. ns not significant; **P < 0.01; one-way ANOVA with Tukey’s post hoc test. Bars represent the mean ± SEM. e–i Representative images of autophagy-related proteins (Beclin1, LC3B, and P62) and KIF5A expression in cultured primary neurons were detected by western blot (n = 3). Quantitative analysis was performed. ns, not significant; *P < 0.05; **P < 0.01 versus WT group; #, P < 0.05; ##, P < 0.01 versus WT + IMQ group; &P < 0.05; &&P < 0.01 versus Tlr7-/- +IMQ group. Two-way ANOVA with Dunnett’s post hoc test. Bars represent the mean ± SEM.