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