Figure 7A Representative immunoblots for TMEM175 expression levels in the midbrains of mice of different ages (W: weeks; M: months). B Statistics of the relative TMEM175 protein levels normalized to GAPDH in (A). Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.D TMEM175 mRNA levels in the midbrains of WT or TMEM175 KO mice were detected by RT-qPCR using SYBR Green PCR Master Mix. Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.E The knockout of TMEM175 in mouse was verified by Western blot of brain tissues.F Representative TMEM175 basal lysosomal currents recorded in macrophages of WT or TMEM175-KO mice. Ψ is lysosomal membrane potential (deﬁned as Vcytosol - Vlumen). The currents were elicited with ramp voltage protocols (-100 to +100 mV in 1 s, Vh = 0 mV). G Statistics of the current amplitudes of (F). Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.H, Quantification of rears in the cylinder (H) pre- and post-MPTP treatment. Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.I Quantification of the latency to fall from a wire hang apparatus (I) pre- and post-MPTP treatment. Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.J Swing Duration CV (CV%) (Left panel) and Ataxia Coefficient in Gait tests 7 days post-MPTP. Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.K Latency to fall in rotarod tests 7 days post-MPTP. Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.L Immunoblots of the TH levels post-MPTP treatment in WT and TMEM175 KO mouse midbrains. M Quantification of the relative TH levels normalized to GAPDH post-MPTP. Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.N TH immunofluorescent labeling on substantia nigra slices of WT or TMEM175-KO mice post-MPTP treatment. Scale bars = 100 μm. O Statistics of the TH-positive neurons in (N). Data information: The data are presented as the mean ± SEM. Statistical significance was analyzed with two-sided student's t-tests, and is indicated with NS for not significant (p > 0.05), * for p < 0.05, ** for p < 0.01, and *** for p < 0.001. n value means the number of biological replicates made for each data point.