Journal: Developmental cell

Article Title: Lysosomal Regulation of Inter-mitochondrial Contact Fate and Motility in Charcot-Marie-Tooth Type 2

doi: 10.1016/j.devcel.2019.05.033

Figure Lengend Snippet: (A) Confocal time-lapse images of inter-mitochondrial contact (M-M) formation (white arrow) and subsequent untethering (yellow arrow) temporally coupled to mitochondria-lysosome (M-L) contact formation and untethering in live HeLa cells (OMM label mApple-TOM20, lysosome/late endosome label LAMP1-mGFP). Scale bar, 0.5µm.

Article Snippet: The following plasmids were obtained from Addgene: LAMP1-mGFP was a gift from Esteban Dell’Angelica (Addgene #34831) ( Falcon-Perez et al., 2005 ), mito-BFP and mCh-Drp1 were gifts from Gia Voeltz (Addgene #49151, #49152) ( Friedman et al., 2011 ), Snap-Omp25 was a gift from David Sabatini (Addgene # 69599) ( Katajisto et al., 2015 ), mito-PAGFP was a gift from Richard Youle (Addgene # 23348) ( Karbowski et al., 2004 ), EGFP-RAB7A WT was a gift from Qing Zhong (Addgene #28047) ( Sun et al., 2010 ), mApple-TOMM20-N-10, mEmerald-TOMM20-C-10, DsRed2-Mito-7, and mCherry-ER-3 were gifts from Michael Davidson (Addgene #54955, #54281, #55838, #55041), and Mfn1-Myc, Mfn1(K88T)-10xmyc, Mfn2-myc and Mfn2 K109A-myc were gifts from David Chan (Addgene #23212, #26050, #23213, #26051) ( Chen et al., 2003 ).

Techniques:

Journal: Developmental cell

Article Title: Lysosomal Regulation of Inter-mitochondrial Contact Fate and Motility in Charcot-Marie-Tooth Type 2

doi: 10.1016/j.devcel.2019.05.033

Figure Lengend Snippet: (A–C) Confocal time lapse images over 90s and quantification of inter-mitochondrial contact (white arrows) with increased tethered duration despite the presence of adjacent mitochondria-lysosome contacts (yellow arrows) in cells expressing mutant RAB7 (Q67L) compared to wild-type RAB7 (WT) (OMM label mApple-TOM20, lysosome/late endosome label LAMP1-mGFP (Control), RAB7(WT)-GFP or RAB7(Q67L)-GFP). Scale bar, 0.5µm. (n = 20 events from 15 cells (Control), n = 41 events from 8 cells (RAB7 (WT)), n = 55 events from 11 cells (RAB7 (Q67L))).

Article Snippet: The following plasmids were obtained from Addgene: LAMP1-mGFP was a gift from Esteban Dell’Angelica (Addgene #34831) ( Falcon-Perez et al., 2005 ), mito-BFP and mCh-Drp1 were gifts from Gia Voeltz (Addgene #49151, #49152) ( Friedman et al., 2011 ), Snap-Omp25 was a gift from David Sabatini (Addgene # 69599) ( Katajisto et al., 2015 ), mito-PAGFP was a gift from Richard Youle (Addgene # 23348) ( Karbowski et al., 2004 ), EGFP-RAB7A WT was a gift from Qing Zhong (Addgene #28047) ( Sun et al., 2010 ), mApple-TOMM20-N-10, mEmerald-TOMM20-C-10, DsRed2-Mito-7, and mCherry-ER-3 were gifts from Michael Davidson (Addgene #54955, #54281, #55838, #55041), and Mfn1-Myc, Mfn1(K88T)-10xmyc, Mfn2-myc and Mfn2 K109A-myc were gifts from David Chan (Addgene #23212, #26050, #23213, #26051) ( Chen et al., 2003 ).

Techniques: Expressing, Mutagenesis

Journal: Developmental cell

Article Title: Lysosomal Regulation of Inter-mitochondrial Contact Fate and Motility in Charcot-Marie-Tooth Type 2

doi: 10.1016/j.devcel.2019.05.033

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The following plasmids were obtained from Addgene: LAMP1-mGFP was a gift from Esteban Dell’Angelica (Addgene #34831) ( Falcon-Perez et al., 2005 ), mito-BFP and mCh-Drp1 were gifts from Gia Voeltz (Addgene #49151, #49152) ( Friedman et al., 2011 ), Snap-Omp25 was a gift from David Sabatini (Addgene # 69599) ( Katajisto et al., 2015 ), mito-PAGFP was a gift from Richard Youle (Addgene # 23348) ( Karbowski et al., 2004 ), EGFP-RAB7A WT was a gift from Qing Zhong (Addgene #28047) ( Sun et al., 2010 ), mApple-TOMM20-N-10, mEmerald-TOMM20-C-10, DsRed2-Mito-7, and mCherry-ER-3 were gifts from Michael Davidson (Addgene #54955, #54281, #55838, #55041), and Mfn1-Myc, Mfn1(K88T)-10xmyc, Mfn2-myc and Mfn2 K109A-myc were gifts from David Chan (Addgene #23212, #26050, #23213, #26051) ( Chen et al., 2003 ).

Techniques: Recombinant, Plasmid Preparation, Software, Microscopy, Transmission Assay

Journal: The Journal of Neuroscience

Article Title: CLC Anion/Proton Exchangers Regulate Secretory Vesicle Filling and Granule Exocytosis in Chromaffin Cells

doi: 10.1523/jneurosci.2439-21.2022

Figure Lengend Snippet: Figure 6. ClC-3c does not localize to late endosomes. A, Representative confocal image of chromaffin cells coex- pressing ClC-3c-eGFP together with the late endosomal marker mRFP-Rab7. Inset, Areas outlined in white boxes; dashed line within the image represents a linear section of the image analyzed in (B). Scale bars: 5 mm; insets, 1 mm. B, Line scan analysis, dashed line in (A) illustrating that ClC-3c exhibits a different subcellular distribution than Rab7. C, Mander’s coefficient analysis for ClC-3c/Rab7 colocalization experiments. ClC-3c positive puncta do not colocalize with Rab7. ClC-3c/Rab7, n = 13. Data are collected from three independent transfected cultures and represented as mean 6 SEM; n denotes the number of analyzed cells.

Article Snippet: Lentiviral transduction was used to express ClC-3b and ClC-3c (Guzman et al., 2015) in combination with fluorescent markers such as Lamp1 (which was a gift from Walther Mothes, plasmid #1817, Addgene; http://www.addgene.org/1817; RRID:Addgene_1817; Sherer et al., 2003); Rab7, Rab11 (a gift from Richard Pagano, plasmid #12605 and #12674, Addgene; Choudhury et al., 2002); TfR (a gift from Gary Banker, plasmid #45060 Addgene; Burack et al., 2000); and VAMP3 and VAMP4 (a gift from Thierry Galli, plasmid #42310 and #42313, Addgene; Galli et al., 1998).

Techniques: Marker, Transfection