Fig. 3

Vps13^−/−Lyn^−/− mice show amelioration of hematologic phenotype and improvement of autophagy and neuroinflammation in basal ganglia. a Quantitation of acanthocytes by brightfield microscopic analysis of peripheral blood smears from Vps13a^−/− and Vps13a^−/−Lyn^−/− mice. Data from 50 visual fields was collected by two blinded researchers. Results are means ± SEM n = 6; °p < 0.002 versus Vps13a^−/− by t-test b Left panel. Red cell distribution histograms generated for red blood cell volume (RBC Volume) and cell haemoglobin concentration (RBC-HC) of RBCs from wild-type (WT) control, Vps13a^−/−, Vps13a^−/−Lyn^−/− and Vps13a^−/−Lyn^−/+ mice. One representative experiment of six with similar results is shown.The blue circle indicates the presence of a subpopulation of dense red cells containing acanthocytes, as described in human patients (Lupo et al. [42]). c Western blot (Wb) analysis of Ulk1 (Atg1) and Rab 5 from red cell cytosolic fractions of wild-type (WT), Vps13a^−/− and Vps13a^−/−Lyn^−/− mice. Catalase was used as protein loading control. Densitometric analyses of the immunoblot bands similar to those shown are presented at right. Data are means ± SEM (n = 6; *P < 0.02 vs. WT; °P < 0.05 vs. Vps13a^−/− by two-way-ANOVA/Bonferroni’s multiple comparison test). d Western blot (Wb) analysis of Beclin-1, Vps34 and p62 in isolated basal ganglia of wild-type (WT), Vps13a^−/− and Vps13a^−/−Lyn^−/− mice. GAPDH served as protein loading control. Densitometric analyses of the immunoblot bands similar to those shown are presented at right. Data are means ± SEM (n = 6; *p < 0.02 vs. WT; °p < 0.02 compared to Vps13a^−/− mice by two-way-ANOVA/Bonferroni’s multiple comparison test). e Western blot (Wb) analysis of K48-ubiquitinated proteins in basal ganglia isolated from wild-type (WT), Vps13a^−/− and Vps13a^−/−Lyn^−/− mice. GAPDH served as protein loading control. Densitometric analyses of the immunoblot bands similar to those shown are presented at right. Data are means ± SEM (n = 6; *p < 0.02 vs. WT; °p < 0.02 compared to Vps13a^−/− mice by two-way-ANOVA/Bonferroni’s multiple comparison test). f Western blot (Wb) analysis of phospho-NF-kB p65 (P-NF-kB), NF-kB in isolated basal ganglia of wild-type (WT), Vps13a^−/− and Vps13a^−/−Lyn^−/− mice. GAPDH served as protein loading control. Densitometric analyses of the immunoblot bands similar to those shown are presented at right. Data are means ± SEM (n = 6; *p < 0.02 vs. WT; °p < 0.02 compared to Vps13a^−/− mice by two-way-ANOVA/Bonferroni’s multiple comparison test)