vsmc line a10  (ATCC)

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A inhibits HG-induced proliferation of A10 cells (A) Structural formula of Gingerenone A (Gin A). (B) Effect of high glucose (HG) on the proliferation of A10 cells. A10 cells were incubated with 10-, 15-, 30-, or 50-mM glucose for 24 h. Cell proliferation was assessed by MTT assay (n = 8, * P < 0.05 vs. control). (C,D) Effect of Gin A on the proliferation of A10 cells. A10 cells were cultured for 24 h in normal medium or HG (30 mM) with Gin A at 0.1, 1, 10 or 100 μM. Cell proliferation was evaluated by MTT assay (C) , and automated cell counting (D) (n = 8, * P < 0.05 vs. control). (E) Representative PCNA immunofluorescence images of A10 cells cultured under control, HG (30 mM) and HG plus Gin A (10 μM) conditions for 24 h. Staining was repeated in three independent experiments with similar results; images are shown for qualitative illustration only and were not subjected to statistical analysis. (F) PCNA protein levels in A10 cells after 24 h of treatment with control medium, Gin A (10 μM), HG (30 mM) or HG plus Gin A (10 μM) for 24 h. Representative western blots and quantification of PCNA expression quantification of PCNA expression (normalized to H3) are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Incubation, MTT Assay, Control, Cell Culture, Cell Counting, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A reduces HG-induced migration of A10 cells (A) Representative Transwell images showing A10 cell migration after 24 h of culture under control conditions, Gin A (10 μM), HG (30 mM) or HG plus Gin A (10 μM). (B) Representative wound-healing images at 0 and 24 h for A10 cells treated as in (A) . (C) Effect of Gin A on the increase in the number of migrating cells induced by HG (n = 10; * P < 0.05 vs. control; # P < 0.05 vs. HG alone). (D) Effect of Gin A on the increased migration distance induced by HG (n = 10; * P < 0.05 vs. control; # P < 0.05 vs. HG alone).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Migration, Control

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A attenuates HG-induced oxidative stress in A10 cells (A) Intracellular ROS levels in A10 cells cultured for 24 h in control medium or HG (30 mM) in the absence or presence of Gin A (10 μM). ROS was evaluated by DHE staining and quantitative statistical data (n = 10; * P < 0.05 vs. control; # P < 0.05 vs. HG alone). (B–D) Oxidative stress-related indicators in A10 cells treated with control medium, Gin A alone (10 μM), HG (30 mM) or HG plus Gin A (10 μM) for 24 h. MDA (B) , T-AOC (C) and SOD activity (D) measured using assay kits (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone). (E) NOX4 protein expression in A10 cells cultured for 24 h under control, HG or HG plus Gin A conditions. Representative western blots and NOX4/GAPDH ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Cell Culture, Control, Staining, Activity Assay, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Gin A activates AMPK and suppresses mTOR/S6K1 signaling in A10 cells (A) Concentration-dependent effects of Gin A on AMPK phosphorylation. A10 cells were treated with Gin A at 0.1, 1, 10 or 100 μM for 24 h, and p-AMPK/AMPK ratios were determined by Western blot. (B) Time course of AMPK activation by Gin A. Cells were exposed to Gin A (10 μM) for 0.5, 1, 3, 6 or 24 h, and p-AMPK/AMPK levels were determined by Western blot. (C) Effects of Gin A on mTOR phosphorylation. A10 cells were treated with Gin A at 0.1, 1, 10 or 100 μM for 24 h, and p-mTOR/mTOR ratios were determined by Western blot. (D) Effects of Gin A on S6K1 phosphorylation. A10 cells were treated with Gin A at 0.1, 1, 10 or 100 μM for 24 h, and p-S6K1/S6K1 ratios determined by Western blot (n = 6; * P < 0.05 vs. control).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Concentration Assay, Phospho-proteomics, Western Blot, Activation Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Gingerenone A attenuates diabetic vascular remodeling through AMPK/mTOR/S6K1 signaling

doi: 10.3389/fphar.2026.1706103

Figure Lengend Snippet: Role of AMPK activation in the Gin A-mediated inhibition of the proliferation and migration of HG-treated A10 cells (A) Effect of the AMPK inhibitor Compound C on Gin A-induced AMPK activation in HG-treated A10 cells. Cells were pre-incubated with Compound C for 1 h and then exposed to HG (30 mM) with or without Gin A (10 μM) for 24 h. Representative blots and p-AMPK/AMPK ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. Gin A alone). (B–D) Effects of Compound C on the Gin A-induced inhibition of A10 cell proliferation and migration. Cell proliferation was determined by the MTT assay (B) . Cell migration was determined by Transwell (C) and wound healing (D) assays (n = 8; * P < 0.05 vs. control; # P < 0.05 vs. HG alone; & P < 0.05 vs. HG + Gin A). (E,F) Effects of Compound C on Gin A-mediated inhibition of mTOR/S6K1 signaling in HG-treated A10 cells. Representative blots and quantification of p-mTOR/mTOR (E) and p-S6K1/S6K1 (F) ratios are shown (n = 6; * P < 0.05 vs. control; # P < 0.05 vs. HG alone; & P < 0.05 vs. HG + Gin A).

Article Snippet: The VSMC line A10 (CRL-1476, ATCC, United States) was cultured in Dulbecco’s modified Eagle’s medium (DMEM, #30-2002, ATCC) supplemented with 10% fetal bovine serum (FBS, #30-2020, ATCC), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Techniques: Activation Assay, Inhibition, Migration, Incubation, Control, MTT Assay

Journal: Journal of Molecular Endocrinology

Article Title: CEBPB/POU2F2 modulates endothelin 1 expression in prehypertensive SHR vascular smooth muscle cells

doi: 10.1530/JME-22-0178

Figure Lengend Snippet: Different transcriptional regulation of the endothelin (ET)-1 promoter enhancer region. (A) Nuclear and cytoplasmic proteins were isolated from vascular smooth muscle cells (VSMCs) of spontaneously hypertensive rates (SHRs) and Wistar-Kyoto (WKY) rats. Western blot analysis of nuclear protein-poly(ADP ribose) polymerase (PARP) to confirm isolation purity. Coomassie blue staining of SDS-PAGE was used as an indicator of protein loading. (B) The DNA of the enhancer region was used as a probe and was incubated with SHR and WKY VSMC nuclear proteins for the EMSA analysis. Competing DNA was a 100-fold concentrated probe. Arrows indicate differences between probe-bound SHR and WKY VSMC nucleoproteins. (C) An RT-PCR analysis of the expressions of Pou1f1 in a pituitary cell line (GH3), smooth muscle cell line (A10), pituitary gland (from SHRs and WKY rats), and VSMCs (from SHRs and WKY rats). (D) Pou2f2 proteins were incubated with POU2F2 consensus sequence probes and enhancer region probes, and an EMSA was used to analyze protein–DNA interactions. Arrows indicate specific binding sites after mixing with competitor DNA of the POU2F2 consensus sequence. Arrowheads indicate specific binding sites after mixing with competitor DNA of the enhanced region sequence. (E) The POU2F2 and CEBPB consensus sequences of DNA sequences indicated on the bottom line were mutated in the 1309prET1 construct and co-transfected with Renilla reporter plasmids in SHR or WKY VSMCs. Statistical data are shown as the mean ± s.e.m. , * P < 0.05, ** P < 0.01. Student's t -test from three independent experiments. Smooth muscle cells were collected from the aortas of three SHR or WKY rats, and each data set consists of three experiments. *vs 1309prET1 in SHR rats. A full color version of this figure is available at https://doi.org/10.1530/JME-22-0178 .

Article Snippet: The rat VSMC line (A10) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Isolation, Western Blot, Staining, SDS Page, Incubation, Reverse Transcription Polymerase Chain Reaction, Sequencing, Binding Assay, Construct, Transfection

Journal: Journal of Molecular Endocrinology

Article Title: CEBPB/POU2F2 modulates endothelin 1 expression in prehypertensive SHR vascular smooth muscle cells

doi: 10.1530/JME-22-0178

Figure Lengend Snippet: Overexpression of POU2F2 and CEBPB affects endogenous endothelin (ET)-1 expression. Total RNA or culture medium from Wistar-Kyoto (WKY) vascular smooth muscle cells (VSMCs) was collected from cells transfected with pcDNA3.1A (3.1A), POU2F2, CEBPB, and POU2F2 + CEBPB expression vectors. (A) Edn1 mRNA expression was examined by a qPCR. Data shown were normalized to the expression of the α-actin reference gene, and the expression level with 3.1A empty vector transfection was set to 1. (B) EDN1 ELISA for measurement of EDN1 secretion in VSMC culture medium. * P < 0.01. *vs 3.1A empty vector transfection. (C and D) Cells were infected with lentivirus-expressing POU2f2 shRNAs (sh- Pou2f2 -1 and sh- Pou2f2 -2) for 24 h. As a negative control, cells were infected with a luciferase shRNA-containing lentivirus (sh-Luc). POU2F2 shRNA infection was confirmed by an RT-qPCR (C) and western blotting (D). Smooth muscle cells were collected from the aortas of three SHR or WKY rats, and each data set consists of three experiments.

Article Snippet: The rat VSMC line (A10) was obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Over Expression, Expressing, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Infection, Negative Control, Luciferase, shRNA, Quantitative RT-PCR, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Nogo-B Receptor Directs Mitochondria-Associated Membranes to Regulate Vascular Smooth Muscle Cell Proliferation

doi: 10.3390/ijms20092319

Figure Lengend Snippet: NgBR expression is downregulated in the pulmonary artery of HPH rat model and in vascular smooth muscle cells (VSMCs) exposed to hypoxia. Rats were housed in hypobaric hypoxia (10.0% O 2 ) for 45 days, and then exposed to reoxygenation at different times. Pulmonary vascular remodeling was evaluated by measuring the ( A ) mPAP and ( B ) weight ratio of RV/ (LV + S). ( C , D ) Histological images of distal pulmonary arteries (arrows)stained with hematoxylin and eosin. Scale bar = 100 µm. The percent medial wall thickness was analyzed using an Olympus IX-70 microscope. n = 44–50 pulmonary arteries (35–100 µm) from three animals/group. ( E ) The small pulmonary arteries (30–140 µm) were co-stained with α-smooth muscle actin (α-SMA; green) and NgBR (red). Nuclei were stained with DAPI (blue). Results were calculated as a relative arbitrary immunofluorescence unit (AFU) using FV10-ASW3.1 software. n = 38–54 pulmonary arteries from three animals/group. Scale bar = 30 µm. The VSMC cell line, A10, was exposed to 4% O 2 for 48 h, followed by exposure to 21% O 2 for 24 h. ( F , G ) Cell proliferation was evaluated by determining PCNA expression. n = 4. Apoptosis was detected by analyzing cleaved-capase-3 expression. n = 3. ** p < 0.01 H48 versus N48, R24 versus N72. ( H ) NgBR expression was normalized to β-actin. n = 3. * p < 0.05.

Article Snippet: The VSMC cell line A10 was purchased from the American Type Culture Collection and cultured in high-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Expressing, Staining, Microscopy, Immunofluorescence, Software

Journal: International Journal of Molecular Sciences

Article Title: Nogo-B Receptor Directs Mitochondria-Associated Membranes to Regulate Vascular Smooth Muscle Cell Proliferation

doi: 10.3390/ijms20092319

Figure Lengend Snippet: A schematic diagram showing how NgBR is involved in MAM-regulated VSMC proliferation. Downregulation of NgBR disrupts the mitochondria-ER unit and increases MAM-associated pAkt-IP 3 R3 signaling, resulting in the suppression of mitochondrial function. Consequently, mitochondria-induced HIF-1α signaling is activated to stimulate cell proliferation.

Article Snippet: The VSMC cell line A10 was purchased from the American Type Culture Collection and cultured in high-glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques:

Journal: Human Molecular Genetics

Article Title: The biological impact of blood pressure-associated genetic variants in the natriuretic peptide receptor C gene on human vascular smooth muscle

doi: 10.1093/hmg/ddx375

Figure Lengend Snippet: The BP-elevating allele at NPR3 locus enhances Ang II-induced human VSMCs contraction. ‘E’ indicates the BP-elevating allele, ‘L’ denotes the BP-lowering allele. ( A ) Representative images of VSMCs of the three different genotypes at baseline (before stimulation) and 30 min after treatment with 100 nmol/l Ang II, respectively. White dash lines show the original size of cells at baseline, and the black ones outline the sizes at 30 min in the merged panel. Scale bars = 50 µm. ( B ) Differences in 100 nmol/l Ang II stimulated VSMC contraction of different genotypes at 15, 30, 45, 60 min. Mean ± SEM, * P < 0.05.

Article Snippet: The VSMCs and ECs were used in all experiments in this study, except the luciferase reporter gene assays in which a rat thoracic aorta VSMC line (A10) (ATCC, CRL-1476) was employed.

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Reduction of blood pressure by store‐operated calcium channel blockers

doi: 10.1111/jcmm.12684

Figure Lengend Snippet: The effect of SK or Gd 3+ on cell proliferation in the presence or absence of LPA . Prior to treatment, the cells were cultured in serum free medium for 20 hrs, then different concentrations of SK were added to different wells and 10 min. later, LPA was added to all the wells except for control (Con) group. After continuing culture for 24 hrs, the cell number was counted. * P < 0.05 compared to control value; # P < 0.05 compared to the group with LPA but no SK or Gd 3+ ; n = 6.

Article Snippet: DMEM and foetal bovine serum (FBS) were obtained from Invitrogen (Burlington, ON, Canada), whereas the A10 VSMC line was from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Cell Culture, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Reduction of blood pressure by store‐operated calcium channel blockers

doi: 10.1111/jcmm.12684

Figure Lengend Snippet: The effect of SK or Gd 3+ on DNA synthesis in the presence or absence of LPA . Prior to treatment, the cells were cultured in serum free medium for 20 hrs, then different concentration of SK or Gd 3+ were added to different wells and 10 min. later, LPA was added to all the wells except for control (Con) group. After incubation for 4 hrs, 3 H‐thymidine was added, then reaction was terminated 20 hrs later. * P < 0.05 compared to control value; # P < 0.05 compared to the group with LPA but no SK or Gd 3+ ; n = 6.

Article Snippet: DMEM and foetal bovine serum (FBS) were obtained from Invitrogen (Burlington, ON, Canada), whereas the A10 VSMC line was from the American Type Culture Collection (Manassas, VA, USA).

Techniques: DNA Synthesis, Cell Culture, Concentration Assay, Control, Incubation