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Developmental Studies Hybridoma Bank anti lamp2
Anti Lamp2, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 93/100, based on 201 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti lamp2/product/Developmental Studies Hybridoma Bank
Average 93 stars, based on 201 article reviews

anti lamp2 - by Bioz Stars, 2026-06

93/100 stars

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Agrimol B blocks autophagic flux in PDAC cells. (A, B) Western blot analysis of P62 and CTSD in PANC-1 and AsPC-1 cells treated with Agrimol B for 24 h. (C, E, F) Immunofluorescence analysis of RFP-GFP-LC3 after PANC-1 and AsPC-1 cells were transfected with RFP-GFP-LC3 for 48 h, followed by treatment with or without Agrimol B for another 24 h. Scale bars, 10 μm. (D, G-L) Immunofluorescence analysis of the colocalization of endogenous LC3 with LAMP2 after treatment with Agrimol B or rapamycin for 24 h in PANC-1 and AsPC-1 cells. Scale bars, 10 μm. (M-O) Immunofluorescence analysis of LC3 in PDAC cells treated with or without Agrimol B in the presence or absence of HCQ. Scale bars, 10 μm.

Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Agrimol B blocks autophagic flux in PDAC cells. (A, B) Western blot analysis of P62 and CTSD in PANC-1 and AsPC-1 cells treated with Agrimol B for 24 h. (C, E, F) Immunofluorescence analysis of RFP-GFP-LC3 after PANC-1 and AsPC-1 cells were transfected with RFP-GFP-LC3 for 48 h, followed by treatment with or without Agrimol B for another 24 h. Scale bars, 10 μm. (D, G-L) Immunofluorescence analysis of the colocalization of endogenous LC3 with LAMP2 after treatment with Agrimol B or rapamycin for 24 h in PANC-1 and AsPC-1 cells. Scale bars, 10 μm. (M-O) Immunofluorescence analysis of LC3 in PDAC cells treated with or without Agrimol B in the presence or absence of HCQ. Scale bars, 10 μm.

Article Snippet: An anti-microtubule-associated protein light chain 3 (LC3) antibody (NB100-2220) was purchased from Novus, while anti-MTERF3 (EM1701-29) and lysosomal associated membrane protein 2 (LAMP2) (M1603-5) antibodies were purchased from HuaBio.

Techniques: Western Blot, Immunofluorescence, Transfection

( A ) Confocal microscopy of endogenous BMP (yellow) and LAMP2 (magenta) immunofluorescence in wild-type (WT) and R1441G LRRK2 MEFs. Scale bar: 20 µm. Quantification of vesicular BMP intensity ( B ) and LAMP2 relative intensity ( C ) per cell area. Colored dots represent mean value from four independent experiments, and violin plots show the distribution of individual cell data. Significance determined by two-tailed paired t -test **p < 0.01, ***p < 0.001. ( D ) Representative transmission electron microscopy (TEM) images of multivesicular endosomes (MVE) from WT and R1441G LRRK2 MEFs. MVB periphery highlighted in yellow. Scale bar: 250 nm. ( E ) MVE area (µm 2 ) quantification in WT and R1441G LRRK2 mutant cells. Colored dots represent mean values from 3 independent experiments and violin plots show the distribution of individual cell data (35–45 cells/group). ( F ) Quantification of intraluminal vesicles (ILVs) per MVE in WT and R1441G LRRK2 MEF cells. The number of ILVs per MVE is binned in three groups and plotted as a percentage of MVE from the total population of each experiment independently. Data from 3 independent experiments (mean ± SEM). Significance determined by two-tailed unpaired t -test ( E ) and ordinary two-way ANOVA, uncorrected Fisher’s LSD ( F ). *p < 0.05, ****p < 0.0001. Figure 1—source data 1. IF images in panel A. Figure 1—source data 2. TEM images in panel D. Figure 1—source data 3. Plotted values in panels B, C, E, F.

Journal: eLife

Article Title: Extracellular vesicle-mediated release of bis(monoacylglycerol)phosphate is regulated by LRRK2 and glucocerebrosidase activity

doi: 10.7554/eLife.106330

Figure Lengend Snippet: ( A ) Confocal microscopy of endogenous BMP (yellow) and LAMP2 (magenta) immunofluorescence in wild-type (WT) and R1441G LRRK2 MEFs. Scale bar: 20 µm. Quantification of vesicular BMP intensity ( B ) and LAMP2 relative intensity ( C ) per cell area. Colored dots represent mean value from four independent experiments, and violin plots show the distribution of individual cell data. Significance determined by two-tailed paired t -test **p < 0.01, ***p < 0.001. ( D ) Representative transmission electron microscopy (TEM) images of multivesicular endosomes (MVE) from WT and R1441G LRRK2 MEFs. MVB periphery highlighted in yellow. Scale bar: 250 nm. ( E ) MVE area (µm 2 ) quantification in WT and R1441G LRRK2 mutant cells. Colored dots represent mean values from 3 independent experiments and violin plots show the distribution of individual cell data (35–45 cells/group). ( F ) Quantification of intraluminal vesicles (ILVs) per MVE in WT and R1441G LRRK2 MEF cells. The number of ILVs per MVE is binned in three groups and plotted as a percentage of MVE from the total population of each experiment independently. Data from 3 independent experiments (mean ± SEM). Significance determined by two-tailed unpaired t -test ( E ) and ordinary two-way ANOVA, uncorrected Fisher’s LSD ( F ). *p < 0.05, ****p < 0.0001. Figure 1—source data 1. IF images in panel A. Figure 1—source data 2. TEM images in panel D. Figure 1—source data 3. Plotted values in panels B, C, E, F.

Article Snippet: Antibody , anti-mouse LAMP2, clone GL2A7 (Rat monoclonal) , Developmental Studies Hybridoma Bank , GL2A7; RRID: AB_2281134 , IF (1:500) WB (1:1000).

Techniques: Confocal Microscopy, Immunofluorescence, Two Tailed Test, Transmission Assay, Electron Microscopy, Mutagenesis

$( A ) Whole cell lysates (WCL) from wild-type (WT) and R1441G LRRK2 mouse embryonic fibroblast (MEF) cells treated with 200 nM MLi-2 for 24 hr were analyzed by immunoblotting. Representative images of LAMP2, phospho-Rab10, and α-Tubulin levels are shown. Molecular weight marker mobility is shown in kDa. ( B ) Flow cytometry measurement of GCase activity using PFB-FDGlu fluorescent GCase substrate in WT and R1441G LRRK2 mutant MEF cells treated with 300 µM conduritol β-epoxide (CBE) for 24 hr. WCL and isolated EVs from WT and R1441G LRRK2 mutant MEF cells treated with 200 nM MLi-2 ( C ) or 300 µM CBE ( G ) for 48 hr were analyzed by immunoblotting. Representative images of LAMP2, Flotillin-1, and α-Tubulin levels are shown. Molecular weight marker mobility is shown in kDa. Immunoblots for LAMP2 and Flotillin-1 in EV fractions required longer exposure times to visualize clear signals across all conditions. Quantification of LAMP2 and Flotillin-1 levels relative to R1441G LRRK2 MEF cells in WCL ( D , H ) and isolated EVs ( E , F , I, J ). Data from 6 to 8 independent experiments (mean ± SEM). Significance determined by Kruskal–Wallis test followed by an uncorrected Dunn’s post hoc test compared to R1441G LRRK2 control *p < 0.05, **p < 0.01, ***p < 0.001; ns, not significant. Figure 2—source data 1. Uncropped blots. Figure 2—source data 2. Annotated uncropped blots. Figure 2—source data 3. Plotted values in panels D–F, H–J.$

Journal: eLife

Article Title: Extracellular vesicle-mediated release of bis(monoacylglycerol)phosphate is regulated by LRRK2 and glucocerebrosidase activity

doi: 10.7554/eLife.106330

Figure Lengend Snippet: ( A ) Whole cell lysates (WCL) from wild-type (WT) and R1441G LRRK2 mouse embryonic fibroblast (MEF) cells treated with 200 nM MLi-2 for 24 hr were analyzed by immunoblotting. Representative images of LAMP2, phospho-Rab10, and α-Tubulin levels are shown. Molecular weight marker mobility is shown in kDa. ( B ) Flow cytometry measurement of GCase activity using PFB-FDGlu fluorescent GCase substrate in WT and R1441G LRRK2 mutant MEF cells treated with 300 µM conduritol β-epoxide (CBE) for 24 hr. WCL and isolated EVs from WT and R1441G LRRK2 mutant MEF cells treated with 200 nM MLi-2 ( C ) or 300 µM CBE ( G ) for 48 hr were analyzed by immunoblotting. Representative images of LAMP2, Flotillin-1, and α-Tubulin levels are shown. Molecular weight marker mobility is shown in kDa. Immunoblots for LAMP2 and Flotillin-1 in EV fractions required longer exposure times to visualize clear signals across all conditions. Quantification of LAMP2 and Flotillin-1 levels relative to R1441G LRRK2 MEF cells in WCL ( D , H ) and isolated EVs ( E , F , I, J ). Data from 6 to 8 independent experiments (mean ± SEM). Significance determined by Kruskal–Wallis test followed by an uncorrected Dunn’s post hoc test compared to R1441G LRRK2 control *p < 0.05, **p < 0.01, ***p < 0.001; ns, not significant. Figure 2—source data 1. Uncropped blots. Figure 2—source data 2. Annotated uncropped blots. Figure 2—source data 3. Plotted values in panels D–F, H–J.

Article Snippet: Antibody , anti-mouse LAMP2, clone GL2A7 (Rat monoclonal) , Developmental Studies Hybridoma Bank , GL2A7; RRID: AB_2281134 , IF (1:500) WB (1:1000).

Techniques: Western Blot, Molecular Weight, Marker, Flow Cytometry, Activity Assay, Mutagenesis, Isolation, Control

( A–C ) No significant differences in EV release between MLi-2/conduritol β-epoxide (CBE)-treated and untreated wild-type (WT) MEF cells. Quantification of LAMP2 and Flotillin-1 levels relative to WT control MEF cells in whole cell lysates (WCL) ( A ) and isolated EVs ( B–C ). Data from 7 to 8 independent experiments (mean ± SEM). Significance determined by ordinary one-way ANOVA, uncorrected Fisher’s LSD. ( D–E ) Characterization of WT or R1441G LRRK2 MEF-derived purified EVs by nanoparticle tracking analysis (NTA). ( D ) Representative plots of EV size distribution in each indicated condition. ( E ) Yield comparison of MEF-derived EVs from each indicated condition determined by NTA. Each colored dot represents an independent experiment. Figure 2—figure supplement 1—source data 1. Plotted values in panels A–C, E.

Journal: eLife

Article Title: Extracellular vesicle-mediated release of bis(monoacylglycerol)phosphate is regulated by LRRK2 and glucocerebrosidase activity

doi: 10.7554/eLife.106330

Figure Lengend Snippet: ( A–C ) No significant differences in EV release between MLi-2/conduritol β-epoxide (CBE)-treated and untreated wild-type (WT) MEF cells. Quantification of LAMP2 and Flotillin-1 levels relative to WT control MEF cells in whole cell lysates (WCL) ( A ) and isolated EVs ( B–C ). Data from 7 to 8 independent experiments (mean ± SEM). Significance determined by ordinary one-way ANOVA, uncorrected Fisher’s LSD. ( D–E ) Characterization of WT or R1441G LRRK2 MEF-derived purified EVs by nanoparticle tracking analysis (NTA). ( D ) Representative plots of EV size distribution in each indicated condition. ( E ) Yield comparison of MEF-derived EVs from each indicated condition determined by NTA. Each colored dot represents an independent experiment. Figure 2—figure supplement 1—source data 1. Plotted values in panels A–C, E.

Article Snippet: Antibody , anti-mouse LAMP2, clone GL2A7 (Rat monoclonal) , Developmental Studies Hybridoma Bank , GL2A7; RRID: AB_2281134 , IF (1:500) WB (1:1000).

Techniques: Control, Isolation, Derivative Assay, Purification, Comparison

( A ) Whole cell lysates (WCL) and isolated EVs from wild-type (WT) mouse embryonic fibroblast (MEF) cells treated with 10 µM GW4869 or 10 nM bafilomycin-A1 (B-A1) for 24 hr were analyzed by immunoblotting. Representative immunoblots of LAMP2, Flotillin-1, and α-Tubulin are shown. Molecular weight marker mobility is shown in kDa. Ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) determination of EV-associated di-18:1-BMP normalized to protein content from WT MEF cells treated with 10 µM GW4869 ( B ) or 10 nM B-A1 ( C ) for 24 hr. Data shown as fold change relative to WT control MEF cells. Only BMP isoforms that were detected are shown. Data from 6 independent experiments (mean ± SEM). Significance determined by two-tailed unpaired t -test; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Quantitation of BMP isoforms normalized to protein content in WCL from MEF WT cells treated with 10 µM GW4869 ( D ) or 10 nM B-A1 ( E ) for 24 hr. Data shown as fold change relative to WT control MEF cells. Only BMP isoforms that were detected are shown. Data from 3 independent experiments (mean ± SEM). Significance determined by two-tailed unpaired t -test; *p < 0.05, **p < 0.01, ***p < 0.001. Figure 6—source data 1. Uncropped blots. Figure 6—source data 2. Annotated uncropped blots. Figure 6—source data 3. Plotted values in panels B–E.

Journal: eLife

Article Title: Extracellular vesicle-mediated release of bis(monoacylglycerol)phosphate is regulated by LRRK2 and glucocerebrosidase activity

doi: 10.7554/eLife.106330

Figure Lengend Snippet: ( A ) Whole cell lysates (WCL) and isolated EVs from wild-type (WT) mouse embryonic fibroblast (MEF) cells treated with 10 µM GW4869 or 10 nM bafilomycin-A1 (B-A1) for 24 hr were analyzed by immunoblotting. Representative immunoblots of LAMP2, Flotillin-1, and α-Tubulin are shown. Molecular weight marker mobility is shown in kDa. Ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) determination of EV-associated di-18:1-BMP normalized to protein content from WT MEF cells treated with 10 µM GW4869 ( B ) or 10 nM B-A1 ( C ) for 24 hr. Data shown as fold change relative to WT control MEF cells. Only BMP isoforms that were detected are shown. Data from 6 independent experiments (mean ± SEM). Significance determined by two-tailed unpaired t -test; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Quantitation of BMP isoforms normalized to protein content in WCL from MEF WT cells treated with 10 µM GW4869 ( D ) or 10 nM B-A1 ( E ) for 24 hr. Data shown as fold change relative to WT control MEF cells. Only BMP isoforms that were detected are shown. Data from 3 independent experiments (mean ± SEM). Significance determined by two-tailed unpaired t -test; *p < 0.05, **p < 0.01, ***p < 0.001. Figure 6—source data 1. Uncropped blots. Figure 6—source data 2. Annotated uncropped blots. Figure 6—source data 3. Plotted values in panels B–E.

Article Snippet: Antibody , anti-mouse LAMP2, clone GL2A7 (Rat monoclonal) , Developmental Studies Hybridoma Bank , GL2A7; RRID: AB_2281134 , IF (1:500) WB (1:1000).

Techniques: Isolation, Western Blot, Molecular Weight, Marker, Liquid Chromatography, Mass Spectrometry, Tandem Mass Spectroscopy, Control, Two Tailed Test, Quantitation Assay

Whole cell lysates (WCL) and isolated EVs from R1441G LRRK2 MEF cells treated with 10 µM GW4869 or 10 nM bafilomycin-A1 for 24 h were analyzed by immunoblotting. Representative images of LAMP2, Flotillin-1, and α-Tubulin levels are shown. Molecular weight marker mobility is shown in kDa. Figure 6—figure supplement 1—source data 1. Uncropped blots. Figure 6—figure supplement 1—source data 2. Annotated uncropped blots.

Journal: eLife

Article Title: Extracellular vesicle-mediated release of bis(monoacylglycerol)phosphate is regulated by LRRK2 and glucocerebrosidase activity

doi: 10.7554/eLife.106330

Figure Lengend Snippet: Whole cell lysates (WCL) and isolated EVs from R1441G LRRK2 MEF cells treated with 10 µM GW4869 or 10 nM bafilomycin-A1 for 24 h were analyzed by immunoblotting. Representative images of LAMP2, Flotillin-1, and α-Tubulin levels are shown. Molecular weight marker mobility is shown in kDa. Figure 6—figure supplement 1—source data 1. Uncropped blots. Figure 6—figure supplement 1—source data 2. Annotated uncropped blots.

Article Snippet: Antibody , anti-mouse LAMP2, clone GL2A7 (Rat monoclonal) , Developmental Studies Hybridoma Bank , GL2A7; RRID: AB_2281134 , IF (1:500) WB (1:1000).

Techniques: Isolation, Western Blot, Molecular Weight, Marker

$a Immunoblot analysis of TBC1D9B -KO (upper panel) and TMEM55B -KO (lower panel) HeLa cells with antibodies against TBC1D9B and TMEM55B. GAPDH serves as a loading control. Two independent clones are shown. n = 3 independent experiments. b Lysosome dispersion in TBC1D9B -KO and TMEM55B -KO cells. Confocal microscopy images of TBC1D9B -KO and TMEM55B -KO cells stained for endogenous LAMP2 (green). Nuclei are stained with DAPI (blue). ns not significant; * = p < 0.05; ** = p < 0.01; wildtype vs. TBC1D9B KO p = 0.0303; wildtype vs. TMEM55B KO p = 0.0073. c Quantification of the number of peripheral lysosomes in wildtype, TBC1D9B -KO, and TMEM55B -KO cells, as determined by OrgaMapper. One-way ANOVA; ns not significant; * = p < 0.05; ** = p < 0.01. Mean ± SEM from n = 3 independent experiments. d Cartoon representation of the TMEM55B/TBC1D9B -KO effects on lysosomal positioning. The KO of TMEM55B or TBC1D9B leads to lysosome dispersion into the cell-periphery. e Quantification of the “percentage of lysosomes in the mobile fraction” of lysotracker red-stained wildtype, TMEM55B -KO, and TBC1D9B -KO cells determined by confocal live-cell imaging microscopy. One-way ANOVA; ns not significant; * = p < 0.05; wildtype vs. TBC1D9B KO p = 0.0225; wildtype vs. TMEM55B KO p = 0.0149; mean ± SEM from n = 1 experiment with wildtype n = 11, TBC1D9B KO n = 12, TMEM55B KO n = 11 measurements. Each measurement represents the mean of all tracks of a cell. f TBC1D9B-HA partially colocalizes with LAMP2 in TBC1D9B KO cells. Confocal images of TBC1D9B KO HeLa cells stained for TBC1D9B-HA (magenta) and endogenous LAMP2 (yellow). g Rescue of defective lysosome position in TBC1D9B -KO cells requires TBC1D9B GAP activity. Immunofluorescence microscopy for LAMP2 (green) of wildtype and untransfected TBC1D9B -KO cells or TBC1D9B -KO cells re-expressing wildtype TBC1D9B-HA (magenta) or a GAP-defective mutant (TBC1D9B RYQ/AAA -HA). Nuclei are stained with DAPI (blue). h Quantification of the number of peripheral lysosomes determined by OrgaMapper. One-way ANOVA; ns not significant; * = p < 0.05; vs. full length p = 0.0428. Mean ± SEM from n = 3 independent experiments.$

Journal: Nature Communications

Article Title: Control of lysosome function by the GTPase-activating protein TBC1D9B and its binding partner TMEM55B

doi: 10.1038/s41467-026-70345-y

Figure Lengend Snippet: a Immunoblot analysis of TBC1D9B -KO (upper panel) and TMEM55B -KO (lower panel) HeLa cells with antibodies against TBC1D9B and TMEM55B. GAPDH serves as a loading control. Two independent clones are shown. n = 3 independent experiments. b Lysosome dispersion in TBC1D9B -KO and TMEM55B -KO cells. Confocal microscopy images of TBC1D9B -KO and TMEM55B -KO cells stained for endogenous LAMP2 (green). Nuclei are stained with DAPI (blue). ns not significant; * = p < 0.05; ** = p < 0.01; wildtype vs. TBC1D9B KO p = 0.0303; wildtype vs. TMEM55B KO p = 0.0073. c Quantification of the number of peripheral lysosomes in wildtype, TBC1D9B -KO, and TMEM55B -KO cells, as determined by OrgaMapper. One-way ANOVA; ns not significant; * = p < 0.05; ** = p < 0.01. Mean ± SEM from n = 3 independent experiments. d Cartoon representation of the TMEM55B/TBC1D9B -KO effects on lysosomal positioning. The KO of TMEM55B or TBC1D9B leads to lysosome dispersion into the cell-periphery. e Quantification of the “percentage of lysosomes in the mobile fraction” of lysotracker red-stained wildtype, TMEM55B -KO, and TBC1D9B -KO cells determined by confocal live-cell imaging microscopy. One-way ANOVA; ns not significant; * = p < 0.05; wildtype vs. TBC1D9B KO p = 0.0225; wildtype vs. TMEM55B KO p = 0.0149; mean ± SEM from n = 1 experiment with wildtype n = 11, TBC1D9B KO n = 12, TMEM55B KO n = 11 measurements. Each measurement represents the mean of all tracks of a cell. f TBC1D9B-HA partially colocalizes with LAMP2 in TBC1D9B KO cells. Confocal images of TBC1D9B KO HeLa cells stained for TBC1D9B-HA (magenta) and endogenous LAMP2 (yellow). g Rescue of defective lysosome position in TBC1D9B -KO cells requires TBC1D9B GAP activity. Immunofluorescence microscopy for LAMP2 (green) of wildtype and untransfected TBC1D9B -KO cells or TBC1D9B -KO cells re-expressing wildtype TBC1D9B-HA (magenta) or a GAP-defective mutant (TBC1D9B RYQ/AAA -HA). Nuclei are stained with DAPI (blue). h Quantification of the number of peripheral lysosomes determined by OrgaMapper. One-way ANOVA; ns not significant; * = p < 0.05; vs. full length p = 0.0428. Mean ± SEM from n = 3 independent experiments.

Article Snippet: LAMP2 (clone H4B4) , DSHB , Cat# sc-18822 , RRID:AB_626858.

Techniques: Western Blot, Control, Clone Assay, Dispersion, Confocal Microscopy, Staining, Live Cell Imaging, Microscopy, Activity Assay, Immunofluorescence, Expressing, Mutagenesis

a Confocal image of wildtype HeLa cells overexpressing TMEM55B (magenta) or TBC1D9B-HA (magenta) stained for endogenous LAMP2 (green). n = 3 independent experiments. b Cartoon representation of the effects of overexpression of TMEM55B or TBC1D9B on lysosome positioning. The overexpression of TMEM55B or TBC1D9B leads to perinuclear clustering of lysosomes. c Confocal images of wildtype HeLa cells overexpressing TBC1D9B RYQ/AAA- HA stained for HA (magenta) and endogenous LAMP2 (green). n = 3 independent experiments. d Pearson correlation coefficient of TBC1D9B/TBC1D9B RYQ/AAA -HA and LAMP2. Mean ± SD from pooled data of n = 3 independent experiments (TBC1D9B n 1 =, n 2 = 27, n 3 = 30; TBC1D9B-RYQ/AAA n 1 = 28, n 2 = 27, n 3 = 21). e Quantification of the number of peripheral lysosomes in each condition was determined by OrgaMapper. One-way ANOVA (left panel), t-test (right panel); ns not significant; * = p < 0.05; ** = p < 0.01. Mean ± SEM from n = 3 independent experiments; left panel: untransfected vs. full length p = 0.043; right panel; untransfected vs. transfected p = 0.0021.

Journal: Nature Communications

Article Title: Control of lysosome function by the GTPase-activating protein TBC1D9B and its binding partner TMEM55B

doi: 10.1038/s41467-026-70345-y

Figure Lengend Snippet: a Confocal image of wildtype HeLa cells overexpressing TMEM55B (magenta) or TBC1D9B-HA (magenta) stained for endogenous LAMP2 (green). n = 3 independent experiments. b Cartoon representation of the effects of overexpression of TMEM55B or TBC1D9B on lysosome positioning. The overexpression of TMEM55B or TBC1D9B leads to perinuclear clustering of lysosomes. c Confocal images of wildtype HeLa cells overexpressing TBC1D9B RYQ/AAA- HA stained for HA (magenta) and endogenous LAMP2 (green). n = 3 independent experiments. d Pearson correlation coefficient of TBC1D9B/TBC1D9B RYQ/AAA -HA and LAMP2. Mean ± SD from pooled data of n = 3 independent experiments (TBC1D9B n 1 =, n 2 = 27, n 3 = 30; TBC1D9B-RYQ/AAA n 1 = 28, n 2 = 27, n 3 = 21). e Quantification of the number of peripheral lysosomes in each condition was determined by OrgaMapper. One-way ANOVA (left panel), t-test (right panel); ns not significant; * = p < 0.05; ** = p < 0.01. Mean ± SEM from n = 3 independent experiments; left panel: untransfected vs. full length p = 0.043; right panel; untransfected vs. transfected p = 0.0021.

Article Snippet: LAMP2 (clone H4B4) , DSHB , Cat# sc-18822 , RRID:AB_626858.

Techniques: Staining, Over Expression, Transfection

a , b Loss of TBC1D9B or TMEM55B impairs starvation-induced perinuclear clustering of lysosomes. a Confocal images of fed and starved (6 h) wildtype, TBC1D9B -KO, and TMEM55B -KO HeLa cells stained for LAMP2. b Quantification of the number of peripheral lysosomes in the different genotypes and under the different conditions by OrgaMapper. One-way ANOVA; ns not significant; * = p < 0.05; ** = p < 0.01; *** = p < 0.001. Mean ± SEM from n = 4 independent experiments; wildtype fed vs. TBC1D9B KO fed p = 0.03; wildtype fed vs. TMEM55B KO fed p = 0.0013; wildtype starved vs. TBC1D9B KO starved p = 0.029; wildtype starved vs. TMEM55B KO starved p = 0.00047. c , d TBC1D9B and TMEM55B are required for the starvation-triggered induction of cathepsin D activity. c SiR-Lysosome staining of live fed- and starved cells (EBSS starved, 4 h) of the indicated genotypes. d Quantification of Cathepsin D activity in starved cells (relative to the unstarved/fed situation). One-way ANOVA; ns not significant; * = p < 0.05. Mean ± SEM from n = 3 independent experiments. e TBC1D9B depletion reduces autophagic flux in starved cells. (Left) immunoblotting and in-gel-fluorescence detection of total lysates of HeLa cells stably expressing Halo-LC3B upon treatment with the indicated siRNAs. (Right) quantification of Halo-LC3B cleavage. Halo band intensities were normalized by the sum of band intensities determined for Halo-LC3B and Halo. ( n = 3 independent experiments). Mean ± SEM from n = 3 independent experiments. Statistics: two-way ANOVA, followed by Tukey´s multiple comparisons test. (siControl-fed vs. siControl-starved: p = 0.002; siTBC1D9B-fed vs siTBC1D9B-starved: P = 0.2046; siControl-fed vs siTBC1D9B-fed: p = 0.9573; siControl-starved vs siTBC1D9B-starved: p = 0.0357).

Journal: Nature Communications

Article Title: Control of lysosome function by the GTPase-activating protein TBC1D9B and its binding partner TMEM55B

doi: 10.1038/s41467-026-70345-y

Figure Lengend Snippet: a , b Loss of TBC1D9B or TMEM55B impairs starvation-induced perinuclear clustering of lysosomes. a Confocal images of fed and starved (6 h) wildtype, TBC1D9B -KO, and TMEM55B -KO HeLa cells stained for LAMP2. b Quantification of the number of peripheral lysosomes in the different genotypes and under the different conditions by OrgaMapper. One-way ANOVA; ns not significant; * = p < 0.05; ** = p < 0.01; *** = p < 0.001. Mean ± SEM from n = 4 independent experiments; wildtype fed vs. TBC1D9B KO fed p = 0.03; wildtype fed vs. TMEM55B KO fed p = 0.0013; wildtype starved vs. TBC1D9B KO starved p = 0.029; wildtype starved vs. TMEM55B KO starved p = 0.00047. c , d TBC1D9B and TMEM55B are required for the starvation-triggered induction of cathepsin D activity. c SiR-Lysosome staining of live fed- and starved cells (EBSS starved, 4 h) of the indicated genotypes. d Quantification of Cathepsin D activity in starved cells (relative to the unstarved/fed situation). One-way ANOVA; ns not significant; * = p < 0.05. Mean ± SEM from n = 3 independent experiments. e TBC1D9B depletion reduces autophagic flux in starved cells. (Left) immunoblotting and in-gel-fluorescence detection of total lysates of HeLa cells stably expressing Halo-LC3B upon treatment with the indicated siRNAs. (Right) quantification of Halo-LC3B cleavage. Halo band intensities were normalized by the sum of band intensities determined for Halo-LC3B and Halo. ( n = 3 independent experiments). Mean ± SEM from n = 3 independent experiments. Statistics: two-way ANOVA, followed by Tukey´s multiple comparisons test. (siControl-fed vs. siControl-starved: p = 0.002; siTBC1D9B-fed vs siTBC1D9B-starved: P = 0.2046; siControl-fed vs siTBC1D9B-fed: p = 0.9573; siControl-starved vs siTBC1D9B-starved: p = 0.0357).

Article Snippet: LAMP2 (clone H4B4) , DSHB , Cat# sc-18822 , RRID:AB_626858.

Techniques: Staining, Activity Assay, Western Blot, Fluorescence, Stable Transfection, Expressing

a TurboID strategy for the identification of TBC1D9B-interacting proteins in iPSC-derived iNeurons. Created in BioRender. Damme, M. (2026) https://BioRender.com/nh1fa4i . b Untargeted label-free quantitation (LFQ) of the ARL8B proximity interactome in iNeurons. c TBC1D9B specifically associates with active ARL8B-GTP. Immunoprecipitation using GFP-Nanotrap antibodies from lysates of HeLa cells expressing different ARL8B variants (wildtype, Q75L, T34N) and TBC1D9B-HA. Samples were analyzed by immunoblotting for GFP and HA. GFP-transfected cells served as a negative control. n = 3 independent experiments. d In vitro pulldown experiment of recombinant purified TBC1D9B-eGFP incubated with different GST-ARL8B variants (Q75L, T34N) bound to beads. Recombinant GST served as a negative control. GST/GST-fusion proteins were visualized by Coomassie staining (false-color blue) and TBC1D9B-GFP by in-gel GFP fluorescence. Quantification of the relative binding of TBC1D9B-GFP to ARL8B Q75L/ T34N. One-sample t-test; two-sided; p = 0.0355. Mean ± SEM from n = 3 independent experiments. e In vitro pulldown experiment of recombinant purified GST-TMEM55B/GST alone and TBC1D9B-eGFP incubated with recombinant ARL8B lacking amino acids 1–17 (ARL8B delta17). GST-TMEM55B/GST were bound to beads. GST/GST-TMEM55B-GST-fusion protein was visualized by Coomassie staining (false-color blue), TBC1D9B-GFP, and ARL8B delta17 by immunoblot. n = 3 independent experiments. f Confocal images of wildtype HeLa cells overexpressing TBC1D9B-HA alone or together with the indicated eGFP-tagged-ARL8B variants (wildtype, Q75L, T34N; white) stained with antibodies against HA (magenta) and LAMP2 (green). Nuclei are stained with DAPI (blue). A representative experiment from 3 replicates is shown for the immunofluorescence panel. Fluorescence intensity profiles are depicted for the indicated cross-sections. The Pearson correlation coefficient of TBC1D9B-HA and LAMP2 is depicted. Bar graph: One-way ANOVA; mean ± SEM from n = 2 independent experiments; ns not significant; **** = p < 0.0001; TBC1D9B o/e vs. TBC1D9B + ARL8B Q75L o/e p = 2.48 × 10 −11 ; TBC1D9B + ARL8B o/e vs. TBC1D9B + ARL8B Q75L o/e p = 9.6 × 10 −13 ; TBC1D9B + ARL8B T34N o/e vs. TBC1D9B + ARL8B Q75L o/e p = 9.6 × 10 −13 ; TBC1D9B o/e n 1 = 29, n 2 = 32; TBC1D9B + ARL8B o/e n 1 = 47, n 2 = 35; TBC1D9B + ARL8B Q75L o/e n 1 = 51, n 2 = 31; TBC1D9B + ARL8B T34N o/e n 1 = 43, n 2 = 39.

Journal: Nature Communications

Article Title: Control of lysosome function by the GTPase-activating protein TBC1D9B and its binding partner TMEM55B

doi: 10.1038/s41467-026-70345-y

Figure Lengend Snippet: a TurboID strategy for the identification of TBC1D9B-interacting proteins in iPSC-derived iNeurons. Created in BioRender. Damme, M. (2026) https://BioRender.com/nh1fa4i . b Untargeted label-free quantitation (LFQ) of the ARL8B proximity interactome in iNeurons. c TBC1D9B specifically associates with active ARL8B-GTP. Immunoprecipitation using GFP-Nanotrap antibodies from lysates of HeLa cells expressing different ARL8B variants (wildtype, Q75L, T34N) and TBC1D9B-HA. Samples were analyzed by immunoblotting for GFP and HA. GFP-transfected cells served as a negative control. n = 3 independent experiments. d In vitro pulldown experiment of recombinant purified TBC1D9B-eGFP incubated with different GST-ARL8B variants (Q75L, T34N) bound to beads. Recombinant GST served as a negative control. GST/GST-fusion proteins were visualized by Coomassie staining (false-color blue) and TBC1D9B-GFP by in-gel GFP fluorescence. Quantification of the relative binding of TBC1D9B-GFP to ARL8B Q75L/ T34N. One-sample t-test; two-sided; p = 0.0355. Mean ± SEM from n = 3 independent experiments. e In vitro pulldown experiment of recombinant purified GST-TMEM55B/GST alone and TBC1D9B-eGFP incubated with recombinant ARL8B lacking amino acids 1–17 (ARL8B delta17). GST-TMEM55B/GST were bound to beads. GST/GST-TMEM55B-GST-fusion protein was visualized by Coomassie staining (false-color blue), TBC1D9B-GFP, and ARL8B delta17 by immunoblot. n = 3 independent experiments. f Confocal images of wildtype HeLa cells overexpressing TBC1D9B-HA alone or together with the indicated eGFP-tagged-ARL8B variants (wildtype, Q75L, T34N; white) stained with antibodies against HA (magenta) and LAMP2 (green). Nuclei are stained with DAPI (blue). A representative experiment from 3 replicates is shown for the immunofluorescence panel. Fluorescence intensity profiles are depicted for the indicated cross-sections. The Pearson correlation coefficient of TBC1D9B-HA and LAMP2 is depicted. Bar graph: One-way ANOVA; mean ± SEM from n = 2 independent experiments; ns not significant; **** = p < 0.0001; TBC1D9B o/e vs. TBC1D9B + ARL8B Q75L o/e p = 2.48 × 10 −11 ; TBC1D9B + ARL8B o/e vs. TBC1D9B + ARL8B Q75L o/e p = 9.6 × 10 −13 ; TBC1D9B + ARL8B T34N o/e vs. TBC1D9B + ARL8B Q75L o/e p = 9.6 × 10 −13 ; TBC1D9B o/e n 1 = 29, n 2 = 32; TBC1D9B + ARL8B o/e n 1 = 47, n 2 = 35; TBC1D9B + ARL8B Q75L o/e n 1 = 51, n 2 = 31; TBC1D9B + ARL8B T34N o/e n 1 = 43, n 2 = 39.

Article Snippet: LAMP2 (clone H4B4) , DSHB , Cat# sc-18822 , RRID:AB_626858.

Techniques: Derivative Assay, Quantitation Assay, Immunoprecipitation, Expressing, Western Blot, Transfection, Negative Control, In Vitro, Recombinant, Purification, Incubation, Staining, Fluorescence, Binding Assay, Immunofluorescence

a Alphafold3 model for TBC1D9B (orange) bound to ARL8B (ruby-colored) in the presence of GTP (yellow sticks). The residues mutated in the TBC1D9B RYQ/AAA version are shown as spheres and highlighted in red. b Closeup view of theTBC1D9B-(GTP)-ARL8B interface. c GTPase activity of 6 μM GTP-bound GST-ARL8B was measured using the GTPase-Glo assay in the presence of 0.5 μM TBC1D9B or TBC1D9B RYQ/AAA at different incubation times. GTPase-Glo™ detects the remaining GTP upon the conversion to ATP; ATP is quantified by the addition of detection reagent and emitted luminescence. The change of luminescence indicates the amount of hydrolyzed GTP in the reaction. Data are presented as mean values ± SEM, n = 3 independent experiments. Two-way ANOVA, Bonferroni’s multiple comparisons test; GST-ARL8B + TBC1D9B vs. GST-ARL8B, at 30 min, p = 1.005 × 10 −5 ; at 60 min, p < 1.4 × 10 −13 ; at 90 min, p < 1 × 10 −14 . d Confocal images of wildtype, TMEM55B knockout, and TBC1D9B knockout HeLa cells stained for LAMP2 and ARL8B-GFP. Left, Pearson correlation coefficient between LAMP2 and ARL8B. One-way ANOVA; mean ± SEM from n = 1 experiment with WT n = 41, TBC1D9B KO n = 39, TMEM55B KO n = 31; ns not significant; **** = p < 0.0001; Wildtype vs. TBC1D9B KO p = 2.37 × 10 −6 ; Wildtype vs. TMEM55B KO p = 5.47 × 10 −7 .

Journal: Nature Communications

Article Title: Control of lysosome function by the GTPase-activating protein TBC1D9B and its binding partner TMEM55B

doi: 10.1038/s41467-026-70345-y

Figure Lengend Snippet: a Alphafold3 model for TBC1D9B (orange) bound to ARL8B (ruby-colored) in the presence of GTP (yellow sticks). The residues mutated in the TBC1D9B RYQ/AAA version are shown as spheres and highlighted in red. b Closeup view of theTBC1D9B-(GTP)-ARL8B interface. c GTPase activity of 6 μM GTP-bound GST-ARL8B was measured using the GTPase-Glo assay in the presence of 0.5 μM TBC1D9B or TBC1D9B RYQ/AAA at different incubation times. GTPase-Glo™ detects the remaining GTP upon the conversion to ATP; ATP is quantified by the addition of detection reagent and emitted luminescence. The change of luminescence indicates the amount of hydrolyzed GTP in the reaction. Data are presented as mean values ± SEM, n = 3 independent experiments. Two-way ANOVA, Bonferroni’s multiple comparisons test; GST-ARL8B + TBC1D9B vs. GST-ARL8B, at 30 min, p = 1.005 × 10 −5 ; at 60 min, p < 1.4 × 10 −13 ; at 90 min, p < 1 × 10 −14 . d Confocal images of wildtype, TMEM55B knockout, and TBC1D9B knockout HeLa cells stained for LAMP2 and ARL8B-GFP. Left, Pearson correlation coefficient between LAMP2 and ARL8B. One-way ANOVA; mean ± SEM from n = 1 experiment with WT n = 41, TBC1D9B KO n = 39, TMEM55B KO n = 31; ns not significant; **** = p < 0.0001; Wildtype vs. TBC1D9B KO p = 2.37 × 10 −6 ; Wildtype vs. TMEM55B KO p = 5.47 × 10 −7 .

Article Snippet: LAMP2 (clone H4B4) , DSHB , Cat# sc-18822 , RRID:AB_626858.

Techniques: Activity Assay, Glo Assay, Incubation, Knock-Out, Staining

$a Confocal images of wildtype, TMEM55B -KO, and TBC1D9B -KO HeLa cells treated with scrambled siRNA or siRNA against ARL8B and stained for endogenous LAMP2. b The fraction of cells with peripheral lysosomes was quantified using OrgaMapper. One-way ANOVA (Šídák’s multiple comparisons test), ** p < 0.01, *** p < 0.001. wildtype scr vs. wildtype siARL8B p = 0.1093; wildtype scr vs. TBC1D9B KO p = 0.0047; scr wildtype scr vs. TMEM55B KO p = 0.0026; scr TBC1D9B KO scr vs. TBC1D9B KO p = 0.0031; siARL8B TMEM55B KO scr vs. TMEM55B KO siARL8B p = 0.0003; mean ± SEM from n = 3 independent experiments. c Confocal images of wildtype and ARL8A/B double-KO (DKO) cells treated with scrambled siRNA or siRNA against TBC1D9B and stained for LC3. d LC3-signal intensity was quantified and expressed as a fold change of scrambled-treated wildtype cells. One-way ANOVA, ns = not significant, **** p < 0.0001. Mean ± SEM from n = 3 independent experiments. e Schematic representation of the proposed model: TMEM55B binds TBC1D9B, which in turn acts as a GAP for ARL8B to facilitate nucleotide exchange and, thereby, membrane association and effector recruitment. TMEM55B-association putatively recruits TBC1D9B to the lysosomal surface and/ or mediates a conformational change to activate TBC1D9B.$

Journal: Nature Communications

Article Title: Control of lysosome function by the GTPase-activating protein TBC1D9B and its binding partner TMEM55B

doi: 10.1038/s41467-026-70345-y

Figure Lengend Snippet: a Confocal images of wildtype, TMEM55B -KO, and TBC1D9B -KO HeLa cells treated with scrambled siRNA or siRNA against ARL8B and stained for endogenous LAMP2. b The fraction of cells with peripheral lysosomes was quantified using OrgaMapper. One-way ANOVA (Šídák’s multiple comparisons test), ** p < 0.01, *** p < 0.001. wildtype scr vs. wildtype siARL8B p = 0.1093; wildtype scr vs. TBC1D9B KO p = 0.0047; scr wildtype scr vs. TMEM55B KO p = 0.0026; scr TBC1D9B KO scr vs. TBC1D9B KO p = 0.0031; siARL8B TMEM55B KO scr vs. TMEM55B KO siARL8B p = 0.0003; mean ± SEM from n = 3 independent experiments. c Confocal images of wildtype and ARL8A/B double-KO (DKO) cells treated with scrambled siRNA or siRNA against TBC1D9B and stained for LC3. d LC3-signal intensity was quantified and expressed as a fold change of scrambled-treated wildtype cells. One-way ANOVA, ns = not significant, **** p < 0.0001. Mean ± SEM from n = 3 independent experiments. e Schematic representation of the proposed model: TMEM55B binds TBC1D9B, which in turn acts as a GAP for ARL8B to facilitate nucleotide exchange and, thereby, membrane association and effector recruitment. TMEM55B-association putatively recruits TBC1D9B to the lysosomal surface and/ or mediates a conformational change to activate TBC1D9B.

Article Snippet: LAMP2 (clone H4B4) , DSHB , Cat# sc-18822 , RRID:AB_626858.

Techniques: Staining, Membrane

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