Fig. 6

Pue treatment enhanced mitochondrial autophagy in the PD cell model via promoting Mfn2 expression. (A-B) Western blot was executed to monitor the expression of autophagy-related proteins in hPDLCs and mitochondria (Full-length blots/gels are presented in Supplementary Fig. 6A-LC3, Fig. 6A-P62, Fig. 6A-GAPDH, Fig. 6A-Mitochondrial LC3, Fig. 6A-Mitochondrial P62, Fig. 6A-Mitochondrial Parkin, Fig. 6A-Mitochondrial Mfn2, Fig. 6A-COXIV). n = 3. (C-D) The PI3K/AKT/mTOR pathway activity in hPDLCs was scrutinized by Western blot (Full-length blots/gels are presented in Supplementary Fig. 6C-AKT, Fig. 6C-p-AKT, Fig. 6C-PI3K, Fig. 6C-p-PI3K, Fig. 6C-mTOR, Fig. 6C-p-mTOR, Fig. 6C-GAPDH). n = 3. (E-F) TMRE staining combined with flow cytometry analysis was performed for the observation of mitochondrial membrane potential changes in hPDLCs. n = 3. (G-H) MitoSox Red staining combined with flow cytometry analysis was employed to detect mitochondrial ROS level in hPDLCs. n = 3. (I-J) mRFP-eGFP-LC3 assay was adopted to explore LC3 dots in hPDLCs. n = 3. (K-L) Parkin/Tomm20 immunofluorescence staining of hPDLCs was executed to evaluate mitochondrial autophagy. n = 3. * P < 0.05. ** P < 0.01. *** P < 0.001