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. 2019 Jul 19;10:1495. doi: 10.3389/fimmu.2019.01495

Figure 2.

Figure 2

Ataxin-3 associates with the mitochondrial protein MIC60 and regulates expression of the OXPHOS machinery components. (A) Silver stain of immunoprecipitation of ataxin-3 in moDCs, using ataxin-3 antibody, isotype control antibody, or immunoprecipitation beads only. (B) The sequence coverage of MIC60 identified following mass spectrometry analysis of the immunoprecipitation of ataxin-3 in moDCs, with the peptides in red indicating the identified peptides. (C) Immunoblot using antibodies against MIC60 and ataxin-3 of lysates where ataxin-3 has been immunoprecipitated from moDCs either left unstimulated or stimulated for 30 or 60 min with MDP + PAM3CSK4. (D) Immunoblot using antibodies against MIC60 and ataxin-3 of lysates where MIC60 has been immunoprecipitated from moDCs. (E) STED microscopy using antibodies against ataxin-3 and MIC60 in moDCs. (F) Immunoblot using antibodies against ataxin-3 and β-actin in moDCs following transfection with control or ataxin-3 siRNA. (G) RT-qPCR analysis of selected mitochondrial genome encoded transcripts following ataxin-3 depletion by siRNA in moDCs; n = 3, one way ANOVA **p < 0.01, ***p < 0.001, ****p < 0.0001 (H) RT-qPCR analysis of MT-ND1 mRNA expression following ataxin-3 depletion by siRNA in moDCs subsequently left unstimulated or stimulated for 6 h with MDP + PAM3CSK4; n = 3, Student's paired t-test *p < 0.05.