antibodies against rab10  (Proteintech)

Journal: Molecular Neurodegeneration

Article Title: LRRK2 kinase activity regulates Parkinson’s disease-relevant lipids at the lysosome

doi: 10.1186/s13024-025-00880-7

Figure Lengend Snippet: LRRK2 activity regulates glucosylceramide and BMP levels to maintain proper lysosomal function. A ) BMP(22:6/22:6) levels were measured in WT parental A549 cells and two clones of LRRK2 R1441G KI A549 cells; data are shown as mean ± SEM; n = 17 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation. B ) Heatmap showing elevated glycosphingolipids including multiple GlcCer species in whole cell extracts from two clones of LRRK2 R1441G KI A549 cells as compared to parental WT cells. The heatmaps were generated as percent of change by normalizing the average of different groups to the average of the WT group; n = 14 independent experiments. The analytes included had nominal p-values (* p ≤ 0.10) for genotype difference in either clone and were grouped based on lipid class. White in the color scale depicts the WT-vehicle as 100%, red shows an accumulation (capped at 350%), and blue shows a reduction. C ) WT and two clones of LRRK2 R1441G KI A549 cells were treated with vehicle or DNL151 (2µM) for 72 h, and the levels of GlcCer(d18:1/24:1) were measured using LC-MS/MS. Data are shown as mean ± SEM; n = 10 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation. D ) WT and one clonal line of LRRK2 R1441G KI A549 cells were treated with increasing concentrations of DNL151, and the levels of GlcCer(d18:1/24:1) were measured using LC-MS/MS. Data are shown as mean ± SEM; n = 4 independent experiments, and statistical significance was determined using non-linear fit following log transformation. E ) Dose-response curves show the percent inhibition of LRRK2 kinase activity as measured by levels of phosphorylated T73 Rab10 in WT and one clonal line of LRRK2 R1441G A549 cells. Data are shown as mean ± SEM; n = 4 independent experiments, and statistical significance was determined using non-linear fit following log transformation. F ) Representative images of DQ-BSA signals (left panels: black and white, right panels: red) in WT A549 cells (top panel), LRRK2 R1441G KI cells (middle panel) and LRRK2 R1441G KI cells treated with DNL151(bottom panel). Nuclei stained with NucBlue (blue); scale bar = 10 μm. G ) The sum of spot intensities of DQ-BSA signal was quantified per cell in WT and two clones of LRRK2 R1441G KI A549 cells. The DQ-BSA signals were normalized to the median within each experiment and to the WT control. Data are shown as mean ± SEM; n = 3 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation, * p ≤ 0.05, ** p ≤ 0.01. H ) WT and two clones of LRRK2 R1441G KI A549 cells were treated with vehicle or DNL151 (2µM) for 72 h, and lysosomal proteolysis was measured using the DQ-BSA-based assay. The sum spot intensities of DQ-BSA signal were quantified per cell. The DQ-BSA signals were normalized to the median within each experiment and to the WT vehicle control. Data are shown as mean ± SEM; n = 6 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001

Article Snippet: Hs00794658_m1 , RAB10 , FAM-MGB.

Techniques: Activity Assay, Clone Assay, Transformation Assay, Generated, Liquid Chromatography with Mass Spectroscopy, Inhibition, Staining, Control

Journal: Molecular Neurodegeneration

Article Title: LRRK2 kinase activity regulates Parkinson’s disease-relevant lipids at the lysosome

doi: 10.1186/s13024-025-00880-7

Figure Lengend Snippet: LRRK2 regulates GCase activity and levels of BMP and glucosylsphingosine in human iPSC-derived microglia. A ) pT73 Rab10 levels and total LRRK2 were quantified from WT and LRRK2 G2019S KI iMicroglia using MSD-based assays. The ratio of pRab10/LRRK2 levels were quantified, and data are shown as mean ± SEM; n = 4 independent experiments, and statistical significance was determined using Student’s t-test. B and C ) The levels of GlcSph and BMP (20:4/20:4) were measured using LC-MS/MS in cell lysates from WT, LRRK2 G2019S KI and LRRK2 KO iMicroglia. Data are shown as mean ± SEM; n = 6 independent experiments, and statistical significance was determined using one-way ANOVA and Tukey’s multiple comparison. D ) Lysosomal GCase activity was assessed using the LysoFQ-GBA probe in WT cells, WT cells treated with the GCase inhibitor CBE, LRRK2 G2019S KI, and LRRK2 KO iMicroglia. The sum of the spot intensities per cell was quantified; data are shown as mean ± SEM; n = 3 independent experiments, and statistical significance was determined using one-way ANOVA and Tukey’s method for multiple comparisons. ** p ≤ 0.01, **** p ≤ 0.0001

Article Snippet: Hs00794658_m1 , RAB10 , FAM-MGB.

Techniques: Activity Assay, Derivative Assay, Liquid Chromatography with Mass Spectroscopy, Comparison

Journal: Molecular Neurodegeneration

Article Title: LRRK2 kinase activity regulates Parkinson’s disease-relevant lipids at the lysosome

doi: 10.1186/s13024-025-00880-7

Figure Lengend Snippet: GCase is necessary and sufficient to mediate LRRK2’s effects on the levels of GCase substrates and BMP, and GCase activity is regulated by the LRRK2 substrates Rab10 and Rab12. A ) WT and one clonal line of GBA1 KO A549 cells were treated with vehicle or DNL151 (2µM) for 72 h, and the levels of GlcCer species were measured using LC-MS/MS. The sum of all GlcCer species was measured, and data are shown as mean ± SEM; n = 6 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation. B and C ) WT and one clonal line of LRRK2 R1441G KI A549 cells were treated with vehicle or imiglucerase (2µM) for 72 h, and the levels of GlcCer and BMP (20:4/20:4) were measured using LC-MS/MS. Data are shown as mean ± SEM; n = 4 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation. D and E ) WT and LRRK2 G2019S iMicroglia were treated with vehicle or imiglucerase (1µM) for 72 h, and the levels of GlcSph and BMP (22:6/22:6) were measured using LC-MS/MS. Data are shown as mean ± SEM; n = 3 independent experiments, and statistical significance was determined using two-way ANOVA with Sidak’s multiple comparison test. F ) WT and two clones of LRRK2 R1441G KI A549 cells were treated with vehicle, DNL151 (2µM), or imiglucerase (2µM) for 72 h. The sum of spot intensities of the DQ-BSA fluorescence signal was quantified per cell. DQ-BSA signal was normalized to the median within each experiment and to the average of the WT controls across experiments. Data are shown as mean ± SEM; n = 6 independent experiments and statistical significance was determined using two-way ANOVA with Sidak’s multiple comparison test following log transformation. G ) siRNA KD screen of LRRK2 substrate Rabs identifies modifiers of GCase activity in A549 cells. Rab expression was transiently knocked down in WT A549 cells using transfection of pooled targeted siRNAs, and GCase activity was evaluated using the live cell LysoFQ-GBA probe. Data are shown as mean ± SEM; n = 7 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation. H ) Reduced GCase activity was confirmed in RAB10 KO and RAB12 KO A549 cells; Data are shown as mean ± SEM; n = 5 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation. I ) Levels of GlcCer(d18:1/24:0) and BMP(22:6/22:6) were measured in WT, RAB10 KO and RAB12 KO A549 cells using LCMS/MS. Data are shown as mean ± SEM; n = 5 independent experiments, and statistical significance was determined using one-way ANOVA following log transformation; * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001

Article Snippet: Hs00794658_m1 , RAB10 , FAM-MGB.

Techniques: Activity Assay, Liquid Chromatography with Mass Spectroscopy, Transformation Assay, Comparison, Clone Assay, Fluorescence, Expressing, Transfection

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Rab10 plays a protective role in the development of pathological cardiac hypertrophy

doi: 10.1016/j.jmccpl.2025.100494

Figure Lengend Snippet: Rab10 was decreased in cultured cardiomyocytes and mouse hearts in response to hypertrophy stimuli. (A) Western blot assay of Rab10 protein levels in rat neonatal cardiomyocytes after 200 nmol/L Ang II treatment at 0, 1, 6, and 24 h. Quantification of the relative Rab10 level (lower, n = 3, three independent replicates). (B and C) Western blot assay of Rab10 protein levels in the heart tissues from WT mice treated with Ang II infusion ( n = 3, three mice) or TAC operation ( n = 4, four mice). Quantification of the relative Rab10 level (lower). Data are presented as mean ± s.e.m., and n represents number of samples. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In total, 50–100 μg of protein was loaded onto SDS-PAGE gels, transferred to nitrocellulose membranes, and incubated with primary antibodies against Rab10 (Proteintech), β-actin (Sigma), phospho-ERK1/2, ERK1/2, phospho-AKT and AKT (Cell Signaling Technology).

Techniques: Cell Culture, Western Blot

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Rab10 plays a protective role in the development of pathological cardiac hypertrophy

doi: 10.1016/j.jmccpl.2025.100494

Figure Lengend Snippet: Rab10 overexpression inhibited Ang II-induced cardiomyocyte hypertrophy and Ang II-initiated signaling pathway. (A) Western blot analysis of exogenous Rab10 in NRCMs infected with Ad-GFP or Ad-Rab10 with anti-Myc or anti-Rab10 antibody. Quantification of the relative Rab10 level (lower, n = 3, three independent replicates). (B) Representative images of NRCMs infected with Ad-GFP or Ad-Rab10 and then treated with 200 nmol/L Ang II for 24 h. Hypertrophy was assessed by cell surface area measurements ( n = 3, 150 cells counted per experiment; right). (C) qPCR analysis of the ANP mRNA level in NRCMs ( n = 3, three independent replicates). (D) Representative immunoblotting analysis of p-ERK1/2, ERK1/2, p-AKT and AKT in NRCMs infected with Ad-GFP or Ad-Rab10 and treated with 200 nmol/L Ang II ( n = 3, three independent replicates). (E) Quantification of the relative protein levels (right). β-actin was used as a loading control. Data are presented as mean ± s.e.m., and n represents number of samples. * P < 0.05 , ** P < 0.01 versus Ad-GFP or saline; # P < 0.05, ## P < 0.01 versus Ad-GFP plus Ang II.

Article Snippet: In total, 50–100 μg of protein was loaded onto SDS-PAGE gels, transferred to nitrocellulose membranes, and incubated with primary antibodies against Rab10 (Proteintech), β-actin (Sigma), phospho-ERK1/2, ERK1/2, phospho-AKT and AKT (Cell Signaling Technology).

Techniques: Over Expression, Western Blot, Infection, Control, Saline

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Rab10 plays a protective role in the development of pathological cardiac hypertrophy

doi: 10.1016/j.jmccpl.2025.100494

Figure Lengend Snippet: Rab10 knockdown promoted Ang II-induced cardiomyocyte hypertrophy and Ang II-initiated signaling pathway. (A) Western blot analysis of endogenous Rab10 in NRCMs infected with Ad-siControl or Ad-siRab10 with anti-Rab10 antibody. Quantification of the relative Rab10 level (lower, n = 3, three independent replicates). (B) Representative images of NRCMs infected with Ad-siControl or Ad-siRab10 and treated with 200 nmol/L Ang II for 24 h. Quantification of cell surface area ( n = 3, 150 cells counted per experiment; right). (C). qPCR analysis of the ANP mRNA level in NRCMs ( n = 3, three independent replicates). (D). Western blot analysis of Rab10, p-ERK1/2, ERK1/2, p-AKT and AKT in NRCMs infected with Ad-siControl or Ad-siRab10 and treated with 200 nmol/L Ang II ( n = 3, three independent replicates). (E) Qunatification of the relative protein levels. β-actin was used as an internal control. Data are presented as mean ± s.e.m., and n represents number of samples. * P < 0.05, ** P < 0.01 versus Ad-siControl or saline; # P < 0.05, ## P < 0.01 versus Ad-siControl plus Ang II.

Article Snippet: In total, 50–100 μg of protein was loaded onto SDS-PAGE gels, transferred to nitrocellulose membranes, and incubated with primary antibodies against Rab10 (Proteintech), β-actin (Sigma), phospho-ERK1/2, ERK1/2, phospho-AKT and AKT (Cell Signaling Technology).

Techniques: Knockdown, Western Blot, Infection, Control, Saline

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Rab10 plays a protective role in the development of pathological cardiac hypertrophy

doi: 10.1016/j.jmccpl.2025.100494

Figure Lengend Snippet: Cardiac-specific overexpression of Rab10 ameliorated TAC-mediated cardiac remodelling in mice. (A) Representative M-mode echocardiograms of WT and Rab10 TG mice treated with sham or TAC surgery, and measurement of ejection fraction (EF%) and fractional shortening (FS%) (lower, n = 6, six mice). (B) Gross morphology of WT and Rab10 TG hearts. Scale bar 0.5 cm. The ratios of heart weight to body weight (HW/BW) ( n = 6, six mice). (C) Heart section area detection of WGA staining (left panel). Scale bar, 100 μm. Quantitative analysis of the cardiac myocyte area (right panel, n = 6, one slice per mouse, six views per slice, 200 cells counted). (D) qPCR analysis of the ANP mRNA levels ( n = 3, three independent replicates). (E) Representative Masson trichrome staining (left panel) and quantification (right panel) of heart sections ( n = 6, one slice per mouse, six views per slice). Scale bar, 200 μm. (F) qPCR analysis of the collagen I mRNA level in heart tissues ( n = 3, three independent replicates). (G) Western blot analysis of Flag-Rab10, p-ERK1/2, ERK1/2, p-AKT and AKT in heart tissues (left panel) and quantification (right panel) ( n = 3, three independent replicates). Actin was used as an internal control. Data are presented as mean ± s.e.m., and n represents number of samples. * P < 0.05, ** P < 0.01 versus sham; # P < 0.05, ## P < 0.01 versus WT plus TAC.

Article Snippet: In total, 50–100 μg of protein was loaded onto SDS-PAGE gels, transferred to nitrocellulose membranes, and incubated with primary antibodies against Rab10 (Proteintech), β-actin (Sigma), phospho-ERK1/2, ERK1/2, phospho-AKT and AKT (Cell Signaling Technology).

Techniques: Over Expression, Staining, Western Blot, Control

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Rab10 plays a protective role in the development of pathological cardiac hypertrophy

doi: 10.1016/j.jmccpl.2025.100494

Figure Lengend Snippet: Rab10 knockdown aggarated TAC-mediated cardiac hypertrophy in mice WT mice were injected with rAAV9-siControl or rAAV9-siRab10 for 2 weeks and then subjected to sham or TAC operation for additional 2 weeks. (A) Representative M-mode echocardiograms, and measurement of ejection fraction (EF%) and fractional shortening (FS%) ( n = 6, six mice). (B) Representative images of H&E staining. Scale bar, 0.5 cm. The ratios of heart weight to body weight (HW/BW) ( n = 6, six mice). (C) Heart section area detection of WGA staining (left panel). Scale bar, 100 μm. Quantitative analysis of the cardiac myocyte area (right panel, n = 6, one slice per mouse, six views per slice, 200 cells counted). (D) qPCR analysis of the ANF mRNA levels ( n = 3, three independent replicates). (E) Representative Masson trichrome staining (left panel) and quantification of heart sections (right panel) ( n = 6, one slice per mouse, six views per slice). Scale bar, 200 μm. (F) qPCR analysis of the collagen I mRNA levels ( n = 3, three independent replicates). (G) Western blot analysis of Rab10, p-ERK1/2, ERK1/2, p-AKT and AKT in heart tissues (left panel) and quantification (right panel) ( n = 3, three technical replicates). Actin was used as an internal control. Data are presented as mean ± s.e.m., and n represents number of samples. * P < 0.05, ** P < 0.01 versus sham; # P < 0.05, ## P < 0.01 versus rAAV/siControl plus TAC.

Article Snippet: In total, 50–100 μg of protein was loaded onto SDS-PAGE gels, transferred to nitrocellulose membranes, and incubated with primary antibodies against Rab10 (Proteintech), β-actin (Sigma), phospho-ERK1/2, ERK1/2, phospho-AKT and AKT (Cell Signaling Technology).

Techniques: Knockdown, Injection, Staining, Western Blot, Control

Journal: Journal of Molecular and Cellular Cardiology Plus

Article Title: Rab10 plays a protective role in the development of pathological cardiac hypertrophy

doi: 10.1016/j.jmccpl.2025.100494

Figure Lengend Snippet: Rab10 was decreased in cultured cardiomyocytes and mouse hearts in response to hypertrophy stimuli and the downstream target of miR-199a. (A) miR-199a seed sequence base pairing with its conserved target sequence in the Rab10 3’UTR from human, mouse and rat. (B) qPCR analysis of miR-199a in NRCMs and heart tissues in response to Ang II ( n = 3, three independent replicates). (C) a diagram of the Rab10 3’UTR mutant reporter plasmid with point or deletion mutations and luciferase assay in HEK293ET cells with cotransfection of the Rab10 wild-type or mutant 3’UTR reporter plasmid and the miR-199a mimic. (D) Dual-luciferase analysis of HEK293ET cells cotransfected with miR-199a mimic or inhibitor and human Rab10 3’UTR reporter plasmid. (E) Immunoblotting analysis of Rab10 in NRCMs transfected with miR-199a mimic or inhibitor ( n = 3, three independent replicates). Data are presented as mean ± s.e.m., and n represents number of samples. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: In total, 50–100 μg of protein was loaded onto SDS-PAGE gels, transferred to nitrocellulose membranes, and incubated with primary antibodies against Rab10 (Proteintech), β-actin (Sigma), phospho-ERK1/2, ERK1/2, phospho-AKT and AKT (Cell Signaling Technology).

Techniques: Cell Culture, Sequencing, Mutagenesis, Plasmid Preparation, Luciferase, Cotransfection, Western Blot, Transfection

Journal: bioRxiv

Article Title: LRRK2 Kinase Mediates Increased GCase Activity in Microglia in Response to Proinflammatory Stimuli

doi: 10.1101/2025.10.10.681687

Figure Lengend Snippet: Rab10 phosphorylation is altered in LRRK2 variant iMGs, while GCase and α-synuclein levels are not a LRRK2 expression and Rab10 phosphorylation in LRRK2 variant iMGs with or without 6 hr 100 nM MLi-2 treatment as measured by WB. n = 3. b Quantification of WB of pT73 Rab10 normalized to total Rab10, levels normalized to isogenic control. c GCase and α-synucleinprotein levels in LRRK2 variant iMGs with or without 6 hr 100 nM MLi-2 treatment as measured by WB. n = 3. d-e Quantification of WB normalized to isogenic control. d Quantification of GCase. e Quantification of α-synuclein. One Way ANOVA with Bonferroni post-hoc test * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: Primary antibodies used and dilutions are as follows: LRRK2 pS1292 1/200 (Abcam ab203181), LRRK2 pS935 1/500 (Abcam ab133450), total LRRK2 1/500 (Abcam ab133474), Rab10 pT73 1/500 (Abcam ab230261), total Rab10 1/500 (Cell Signaling 8127S), Rab12 pS106 1/1,000 (Abcam ab256765), total Rab12 1/500 (Proteintech 18843-1-AP), GCase 1/1,000 (Abcam ab55080), α-synuclein 1/1,000 (BD Biosciences 610787), LAMP1 1/1,000 (Cell Signaling 9091S), α-actin 1/50,000 (Millipore MAB1501).

Techniques: Phospho-proteomics, Variant Assay, Expressing, Control

Journal: bioRxiv

Article Title: LRRK2 Kinase Mediates Increased GCase Activity in Microglia in Response to Proinflammatory Stimuli

doi: 10.1101/2025.10.10.681687

Figure Lengend Snippet: LRRK2 inhibition leads to Rab10 dephosphorylation but has no effect on GCase activity. a Percent change in PFB-FDGlu fluorescence. n = 4. b Rab10 phosphorylation is decreased to a similar extent by 6 hr 100 nM MLi-2 or 500 nM PF-475 treatment. n = 2 c Mean total lysotracker fluorescence area per cell (px ) at baseline (0 minutes). a & c Repeated Measures One Way ANOVA Tukey post-hoc test * p < 0.05, ** p < 0.01, **** p < 0.0001

Article Snippet: Primary antibodies used and dilutions are as follows: LRRK2 pS1292 1/200 (Abcam ab203181), LRRK2 pS935 1/500 (Abcam ab133450), total LRRK2 1/500 (Abcam ab133474), Rab10 pT73 1/500 (Abcam ab230261), total Rab10 1/500 (Cell Signaling 8127S), Rab12 pS106 1/1,000 (Abcam ab256765), total Rab12 1/500 (Proteintech 18843-1-AP), GCase 1/1,000 (Abcam ab55080), α-synuclein 1/1,000 (BD Biosciences 610787), LAMP1 1/1,000 (Cell Signaling 9091S), α-actin 1/50,000 (Millipore MAB1501).

Techniques: Inhibition, De-Phosphorylation Assay, Activity Assay, Fluorescence, Phospho-proteomics

Journal: bioRxiv

Article Title: LRRK2 Kinase Mediates Increased GCase Activity in Microglia in Response to Proinflammatory Stimuli

doi: 10.1101/2025.10.10.681687

Figure Lengend Snippet: Inflammatory stimulus does not induce LRRK2-dependent changes in GCase protein expression, lysosomal proteolytic capacity, or pH. a Western blot analysis to assess LRRK2 levels and activity and lysosomal content in LWT iMGs with and without 24 hour 20 ng/mL IFNγ and 25 hour 100 nM MLi-2 treatment. n = 3. b-f Quantification of western blot. b Quantification of pS935 normalized to total LRRK2. c Quantification of LRRK2. d Quantification of pT73 normalized to total Rab10. e Quantification of GCase. f Quantification of LAMP1. g Representative images of DQ Red BSA signal in LWT iMGs following IFNγ and MLi-2 treatment 24 hours after dye-loading. h Mean DQ Red BSA fluorescence intensity normalized to mean total lysotracker area (px ) per cell, normalized to DMSO treated control, 24 hours after dye-loading. n = 3. i Representative images of pHLys Green signal in LWT iMGs following IFNγ and MLi-2 treatment. j Mean pHLys Green fluorescence intensity normalized to mean total LysoPrime Deep Red area (px ) per cell, normalized to DMSO treated control. n = 3. b-i One Way ANOVA with Bonferroni post-hoc test * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Primary antibodies used and dilutions are as follows: LRRK2 pS1292 1/200 (Abcam ab203181), LRRK2 pS935 1/500 (Abcam ab133450), total LRRK2 1/500 (Abcam ab133474), Rab10 pT73 1/500 (Abcam ab230261), total Rab10 1/500 (Cell Signaling 8127S), Rab12 pS106 1/1,000 (Abcam ab256765), total Rab12 1/500 (Proteintech 18843-1-AP), GCase 1/1,000 (Abcam ab55080), α-synuclein 1/1,000 (BD Biosciences 610787), LAMP1 1/1,000 (Cell Signaling 9091S), α-actin 1/50,000 (Millipore MAB1501).

Techniques: Expressing, Western Blot, Activity Assay, Fluorescence, Control