Journal: Traffic (Copenhagen, Denmark)

Article Title: Acute GARP Depletion Disrupts Vesicle Transport, Leading to Severe Defects in Sorting, Secretion and O ‐Glycosylation

doi: 10.1111/tra.70003

Figure Lengend Snippet: List of primary and secondary antibodies.

Article Snippet: CD‐M6PR/MPR , Santa Cruz, #sc365196 , Mouse , 1:1000 , —.

Techniques: Transduction

Journal: bioRxiv

Article Title: EMT activates ER-to-Golgi trafficking through upregulation of REEP2 to promote lung cancer progression

doi: 10.1101/2025.11.12.688098

Figure Lengend Snippet: (A) Schematic illustration of the retention using selective hooks (RUSH) system. The cells were co-transfected with the reporter protein (CD-MPR), which is fused to the streptavidin-binding peptide (SBP) and GFP, and a second protein with a C-terminal ER retention signal (Lys-Asp-Glu-Leu; KDEL) fusing with streptavidin as a hook. The interaction of CD-MPR with the hook protein retains CD-MPR in the ER compartment and biotin administration releases the CD-MPR-GFP reporter from the ER and transfers it to the Golgi. (B) Schematic of VSV-G assay. The cells were transfected with vectors that express an EGFP-tagged temperature-sensitive mutant VSV-G (EGFP-VSV-G [ts045]) that is transported from the ER to the plasma membrane via the Golgi. At designated time points after switching cells to temperatures that cause VSV-G accumulation (40°C) and release (32°C) from the ER, cells were fixed and exofacial VSV-G was detected in non-permeabilized cells by staining with anti-VSV-G monoclonal antibody. The VSV-G trafficking to the plasma membrane is determined by the ratio of exofacial (surface) VSV-G fluorescence signal to the EGFP signal intensity. (C) Confocal micrographs of cells co-stained with anti-SEC16A (green) and anti-GM130 (red) antibodies. DAPI (blue). Scale bar: 5 μm. The scatter plots quantify Golgi-localized SEC16A per cell (dot) based on % of total SEC16A that co-localizes with Golgi (GM130 channel) (n = 15 cells per group) in H1299 cells treated with vehicle control (Veh) or 5 µM thapsigargin (Tg) for 1 h. (D) Confocal micrographs of EGFP-VSV-G-transfected cells taken 1 h after transfer to permissive temperature. Scale bar, 5 μm. The scatter plot represents the ratio of surface VSV-G to EGFP-VSV-G in each cell (dot) (n = 15 cells per group) in H1299 cells treated with vehicle control (Veh) or 5 µM thapsigargin (Tg) for 1 h. Results represent means ± SEM. P values were determined using two-tailed Student’s t-test.

Article Snippet: We purchased fetal bovine serum (FBS), live-cell imaging solution, phosphate-buffered saline (PBS), RPMI-1640, Trypsin-EDTA (0.25%), Alexa Fluor-tagged secondary antibodies, paraformaldehyde, bovine serum albumin (BSA), DAPI and Triton X-100, High-Capacity cDNA Reverse Transcription Kit, PowerUpTM SYBRTM Green Master Mix, and protease/phosphatase inhibitor cocktail from Thermo Fisher Scientific; Doxycycline (#D9891) from MilliporeSigma; Bronchial Epithelial Cell Growth Medium BulletKit from Lonza; jetPRIME transfection reagent from Polyplus Transfection; Transwell and Matrigel-coated Boyden chambers from BD Biosciences; Glass-bottom dishes and multiwell plates from MatTek; 10X Cell lysis buffer from Cell Signaling Technologies; 2X cell lysis buffer (#1610737) from Bio-Rad; CCK-8 (#K1018) from APExBIO; RNeasy Plus Mini Kit (#74136), and AllStars Negative Control siRNA (#1027281) from QIAGEN; Pre-miRTM miRNA Precursor Negative Control (miR-NC) (#AM17110); human miR-183a mimics (#PM12303) and human miR-193 mimics (#PM11786) from Thermo Fisher Scientific; human REEP2 siRNA #2 (#J-021201-18), and #4 (J-021201-20) from Horizon; primary antibodies against β-actin (#4967), ZEB1 (#3396) from Cell Signaling Technologies, REEP2 (#Ab191410) from Abcam, SEC16A (#20025-1-AP) from Proteintech, VSV-G (#EB0012) from Kerafast; Alexa Fluor 555 Mouse anti-GM130 (#560066) from BD Biosciences; REEP2-GFP (#RC202507L4) from Origene; SEC24D-mCherry (#32677), Golgi-pmTurquoise2 (#36205), mCherry-Golgi (#55052) and CD-MPR-GFP-RUSH (#202797) from Addgene; 3’-UTR luciferase reporters of REEP2 (#38786081) from Applied Biological Materials; pCI-Neo Renilla luciferase reporter (Basic) (#E1841) and pGL3 Basic Firefly luciferase control reporter (#E1751) from Promega; All-in-One miRNA qRT-PCR Detection Kit 2.0 (#QP115) from GeneCopoeia.

Techniques: Transfection, Binding Assay, Mutagenesis, Clinical Proteomics, Membrane, Staining, Fluorescence, Control, Two Tailed Test

Journal: bioRxiv

Article Title: EMT activates ER-to-Golgi trafficking through upregulation of REEP2 to promote lung cancer progression

doi: 10.1101/2025.11.12.688098

Figure Lengend Snippet: (A) Confocal micrographs of cells co-transfected with CD-MPR-RUSH-GFP (green) and Golgi-pmTurquoise2 (red). Scale bar: 5 μm. The scatter plots quantify Golgi-localized CD-MPR per cell (dot) based on % of total CD-MPR that co-localizes with Golgi at different time points after biotin treatment (40 μM) (n = 15 cells per group) in H1299 cells transfected with control siRNA (siCTL) or REEP2 siRNA (siREEP2). (B) Confocal micrographs of EGFP-VSV-G-transfected cells taken 1 h after transfer to permissive temperature. Scale bar, 5 μm. The scatter plot represents the ratio of surface VSV-G to EGFP-VSV-G in each cell (dot) (n = 15 cells per group) in HCC827 cells co-transfected with empty vector (Vec) or ZEB1 with control siRNA (siCTL) or REEP2 siRNA (siREEP2). Results represent means ± SEM. P values were determined using two-tailed Student’s t-test.

Article Snippet: We purchased fetal bovine serum (FBS), live-cell imaging solution, phosphate-buffered saline (PBS), RPMI-1640, Trypsin-EDTA (0.25%), Alexa Fluor-tagged secondary antibodies, paraformaldehyde, bovine serum albumin (BSA), DAPI and Triton X-100, High-Capacity cDNA Reverse Transcription Kit, PowerUpTM SYBRTM Green Master Mix, and protease/phosphatase inhibitor cocktail from Thermo Fisher Scientific; Doxycycline (#D9891) from MilliporeSigma; Bronchial Epithelial Cell Growth Medium BulletKit from Lonza; jetPRIME transfection reagent from Polyplus Transfection; Transwell and Matrigel-coated Boyden chambers from BD Biosciences; Glass-bottom dishes and multiwell plates from MatTek; 10X Cell lysis buffer from Cell Signaling Technologies; 2X cell lysis buffer (#1610737) from Bio-Rad; CCK-8 (#K1018) from APExBIO; RNeasy Plus Mini Kit (#74136), and AllStars Negative Control siRNA (#1027281) from QIAGEN; Pre-miRTM miRNA Precursor Negative Control (miR-NC) (#AM17110); human miR-183a mimics (#PM12303) and human miR-193 mimics (#PM11786) from Thermo Fisher Scientific; human REEP2 siRNA #2 (#J-021201-18), and #4 (J-021201-20) from Horizon; primary antibodies against β-actin (#4967), ZEB1 (#3396) from Cell Signaling Technologies, REEP2 (#Ab191410) from Abcam, SEC16A (#20025-1-AP) from Proteintech, VSV-G (#EB0012) from Kerafast; Alexa Fluor 555 Mouse anti-GM130 (#560066) from BD Biosciences; REEP2-GFP (#RC202507L4) from Origene; SEC24D-mCherry (#32677), Golgi-pmTurquoise2 (#36205), mCherry-Golgi (#55052) and CD-MPR-GFP-RUSH (#202797) from Addgene; 3’-UTR luciferase reporters of REEP2 (#38786081) from Applied Biological Materials; pCI-Neo Renilla luciferase reporter (Basic) (#E1841) and pGL3 Basic Firefly luciferase control reporter (#E1751) from Promega; All-in-One miRNA qRT-PCR Detection Kit 2.0 (#QP115) from GeneCopoeia.

Techniques: Transfection, Control, Plasmid Preparation, Two Tailed Test

Journal: Cell reports

Article Title: Post-Golgi carriers, not lysosomes, confer lysosomal properties to pre-degradative organelles in normal and dystrophic axons

doi: 10.1016/j.celrep.2021.109034

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT OR RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-human CTSD This study N/A Sheep anti-human CTSD Cataldo et al., 1990 N/A Rabbit anti-mouse CTSD Cataldo et al., 2004 N/A Sheep anti-TGN38 Bio-rad Cat# AHP499G; RRID: AB_2203272 Rabbit anti-CTSD Scripps Laboratories Cat# RC245 Goat anti-CTSB Neuromics Cat# GT15047; RRID: AB_2737184 Rabbit anti-CTSL Athens Research & Technology Cat# 01–12-030112; RRID: AB_11190245 Mouse anti-alpha-Tubulin Sigma Cat# T9026; RRID: AB_477593 Rabbit anti-mTOR Cell Signaling Cat# 2983s; RRID: AB_2105622 Rabbit anti-p-mTOR Cell Signaling Cat# 2974s; RRID: AB_2262884 Rabbit anti-p-ATG14 Cell Signaling Cat# 13155; RRID: AB_2798133 Rabbit anti-TFEB Bethyl Laboratories Cat# A303–673A; RRID: AB_11204751 Rabbit anti-TFE3 Sigma Cat# HPA023881; RRID: AB_1857931 Rabbit anti-ubiquitin Dako Cat# Z0458; RRID: AB_2315524 Rabbit anti-LC3 Novus Biologicals Cat# NB100–2220; RRID: AB_10003146 Murine anti-NeuN Millipore Cat# MAB377MI; RRID: AB_2298772 Rabbit anti-GFAP Sigma Cat# G9269; RRID: AB_477035 Rabbit anti-Iba-1 Waco Chemicals Cat# 019–19741; RRID: AB_839504 Rat anti-LAMP1 DSHB Cat# 1D4B; RRID: AB_2134500 Rabbit anti-IGF2R (CI-MPR) Sigma Cat# HPA011332; RRID: AB_1846276 Sheep anti-mouse M6PR (CD-MPR) LSBio Cat# LS- {"type":"entrez-nucleotide","attrs":{"text":"C94323","term_id":"3218938","term_text":"C94323"}} C94323 ; RRID: AB_1935265 Goat anti-mouse IgG, Alexa Fluor 647-conjugated Thermo Fisher Cat# A-21236; RRID: AB_2535805 Goat anti-mouse IgG, Alexa Fluor 488-conjugated Thermo Fisher Cat# A-11029; RRID: AB_138404 Goat anti-rabbit IgG, Alexa Fluor 568-conjugated Thermo Fisher Cat# A-11036; RRID: AB_10563566 Donkey anti-rat IgG, Alexa Fluor 647-conjugated Jackson ImmunoReseach Cat# 712–605-153; RRID: AB_2340694 Donkey anti-rabbit IgG, Alexa Fluor 546-conjugated Thermo Fisher Cat# A-10040; RRID: AB_2534016 Donkey anti-sheep IgG, Alexa Fluor 488-conjugated Thermo Fisher Cat# A-11015; RRID: AB_141362 Goat anti-rabbit IgG, biotinylated Vector Laboratories Cat# BA-1000; RRID: AB_2313606 Goat anti-mouse IgG, biotinylated Vector Laboratories Cat# BA-2000; RRID: AB_2313581 Donkey anti-rabbit IgG, 6 nm gold-conjugated Electron Microscopy Sciences Cat# 25703 Donkey anti-mouse IgG, 6 nm gold-conjugated Electron Microscopy Sciences Cat# 25813 Donkey anti-sheep IgG, 10 nm gold-conjugated Electron Microscopy Sciences Cat# 25825 Chemicals, peptides, and recombinant proteins Sodium cacodylate buffer Electron Microscopy Sciences Cat# 11652 Permount Electron Microscopy Sciences Cat# 17986 Fluoro-Gel Electron Microscopy Sciences Cat# 17985–10 Vectorstain ABC Kit Vector Laboratories Cat# PK-4000 Vector DAB Peroxidase Substrate Vector Laboratories Cat# SK-4100 M.O.M.

Techniques: Plasmid Preparation, Electron Microscopy, Recombinant, Immunodetection, Fluorescence, Transfection, Expressing, Software

Journal: Cell reports

Article Title: Post-Golgi carriers, not lysosomes, confer lysosomal properties to pre-degradative organelles in normal and dystrophic axons

doi: 10.1016/j.celrep.2021.109034

Figure Lengend Snippet: Time-lapse imaging in axonal segments of neurons expressing LAMP1-YFP co-labeled with LysoTracker (LT)-Red (A–C) or expressing LAMP1-mCherry co-labeled with dextran-Oregon Green 488 (OG488) and dextran-Alexa Fluor 647 (AF647) (D–F).

Article Snippet: ​ REAGENT OR RESOURCE SOURCE IDENTIFIER Antibodies Mouse monoclonal anti-human CTSD This study N/A Sheep anti-human CTSD Cataldo et al., 1990 N/A Rabbit anti-mouse CTSD Cataldo et al., 2004 N/A Sheep anti-TGN38 Bio-rad Cat# AHP499G; RRID: AB_2203272 Rabbit anti-CTSD Scripps Laboratories Cat# RC245 Goat anti-CTSB Neuromics Cat# GT15047; RRID: AB_2737184 Rabbit anti-CTSL Athens Research & Technology Cat# 01–12-030112; RRID: AB_11190245 Mouse anti-alpha-Tubulin Sigma Cat# T9026; RRID: AB_477593 Rabbit anti-mTOR Cell Signaling Cat# 2983s; RRID: AB_2105622 Rabbit anti-p-mTOR Cell Signaling Cat# 2974s; RRID: AB_2262884 Rabbit anti-p-ATG14 Cell Signaling Cat# 13155; RRID: AB_2798133 Rabbit anti-TFEB Bethyl Laboratories Cat# A303–673A; RRID: AB_11204751 Rabbit anti-TFE3 Sigma Cat# HPA023881; RRID: AB_1857931 Rabbit anti-ubiquitin Dako Cat# Z0458; RRID: AB_2315524 Rabbit anti-LC3 Novus Biologicals Cat# NB100–2220; RRID: AB_10003146 Murine anti-NeuN Millipore Cat# MAB377MI; RRID: AB_2298772 Rabbit anti-GFAP Sigma Cat# G9269; RRID: AB_477035 Rabbit anti-Iba-1 Waco Chemicals Cat# 019–19741; RRID: AB_839504 Rat anti-LAMP1 DSHB Cat# 1D4B; RRID: AB_2134500 Rabbit anti-IGF2R (CI-MPR) Sigma Cat# HPA011332; RRID: AB_1846276 Sheep anti-mouse M6PR (CD-MPR) LSBio Cat# LS- {"type":"entrez-nucleotide","attrs":{"text":"C94323","term_id":"3218938","term_text":"C94323"}} C94323 ; RRID: AB_1935265 Goat anti-mouse IgG, Alexa Fluor 647-conjugated Thermo Fisher Cat# A-21236; RRID: AB_2535805 Goat anti-mouse IgG, Alexa Fluor 488-conjugated Thermo Fisher Cat# A-11029; RRID: AB_138404 Goat anti-rabbit IgG, Alexa Fluor 568-conjugated Thermo Fisher Cat# A-11036; RRID: AB_10563566 Donkey anti-rat IgG, Alexa Fluor 647-conjugated Jackson ImmunoReseach Cat# 712–605-153; RRID: AB_2340694 Donkey anti-rabbit IgG, Alexa Fluor 546-conjugated Thermo Fisher Cat# A-10040; RRID: AB_2534016 Donkey anti-sheep IgG, Alexa Fluor 488-conjugated Thermo Fisher Cat# A-11015; RRID: AB_141362 Goat anti-rabbit IgG, biotinylated Vector Laboratories Cat# BA-1000; RRID: AB_2313606 Goat anti-mouse IgG, biotinylated Vector Laboratories Cat# BA-2000; RRID: AB_2313581 Donkey anti-rabbit IgG, 6 nm gold-conjugated Electron Microscopy Sciences Cat# 25703 Donkey anti-mouse IgG, 6 nm gold-conjugated Electron Microscopy Sciences Cat# 25813 Donkey anti-sheep IgG, 10 nm gold-conjugated Electron Microscopy Sciences Cat# 25825 Chemicals, peptides, and recombinant proteins Sodium cacodylate buffer Electron Microscopy Sciences Cat# 11652 Permount Electron Microscopy Sciences Cat# 17986 Fluoro-Gel Electron Microscopy Sciences Cat# 17985–10 Vectorstain ABC Kit Vector Laboratories Cat# PK-4000 Vector DAB Peroxidase Substrate Vector Laboratories Cat# SK-4100 M.O.M.

Techniques: Imaging, Expressing, Labeling