Plamp1 mcherry

The expression plasmids pEGFP-hGalectin-3 (GFP-Gal3) and pmRFP-EGFP-Galectin-3 (ptf-Gal3) were purchased from Addgene [deposited by Dr. Tamotsu Yoshimori (Osaka University, Japan)]. The subcellular localization vectors, pmTurquoise2-ER, pmTurquoise2-Golgi, and pmTurquoise2-Peroxi, in which a cyan fluorescence protein (Turquoise) was fused with ER, Golgi, or peroxisome targeting sequences respectively, were obtained from Addgene [deposited by Dr. Dorus Gadella (University of Amsterdam, Netherlands)]. pEGFP-TFEB was obtained from Addgene [deposited by Dr. Shawn Ferguson (Yale Univeristy, USA)]. pLAMP1-GFP and pEGFP-LC3 were kindly provided by Dr. Peter K. Kim (Toronto University, Canada) and Dr. Noboru Mizhushima (Tokyo University, Japan), respectively. pCMV-lyso-pHluorin (Lyso-pHluorin) was purchased from Addgene [deposited by Dr. Christian Rosenmund (Neuroscience Research Center, Germany)]. pEGFP-Ubiquitin (GFP-Ub) and pLAMP1-mCherry were purchased from Addgene [deposited by Dr. Nico Dantuma (Karolinska Institutet, Sweden) and Amy Pamer (University of Colorado, USA)], respectively. Mitochondrial-YFP (pMito-YFP) plasmid was kindly provided by Dr. Gyesoon Yoon (Ajou University, Korea).

Mito-mRaspberry-7, mito-meGFP, mito-BFP, EBFP2-C1 (cell fill), iRFP-C1 (cell fill), YFP-Parkin, pPBbsr2-4031NES (AMPK FRET biosensor), snap-OMP25 (SNAPmito), iRFP-Rab7, pLAMP1–mCherry, pLV-TetO-hNGN2-eGFP-puro and FudeltaGW-rtTA were acquired from Addgene (55931, 172481, 49151, 54665, 54786, 23955, 105241, 69599, 51613, 45147, 79823 and 19780, respectively). Plasmids encoding shRNA against SYNJ2BP, AMPK α1 and α2, and non-targeting (TRCN0000139049, TRCN0000024000, TRCN0000024046 and TR30021, respectively) were purchased in pLKO from Sigma-Aldrich. PINK1-kinase dead-MS2-PP7, Cox4i-MS2-PP7, Atp5f1b-MS2-PP7, split Venus, shRNA-resistant myc-tagged SYNJ2BP WT, SYNJ2mito WT and VQL/AAA plasmids have previously been described³ . SYNJ2BP mutations S21A and S21E were introduced by site-directed mutagenesis. PINK1–GFP was generated in pHAGE by restriction-free cloning. The lentiviral packaging plasmids pMDLg/pRRE, pRSV-Rev and pMD2.G were acquired from Addgene (12251, 12253 and 12259, respectively). For purification of SYNJ2BP from E. coli, the cytosolic domain of rat SYNJ2BP WT (amino acids 1–110) was inserted into the bacterial expression vector pET19b and C-terminally tagged with 6xHis-tag.