mouse smooth muscle protein 22 α  (Proteintech)

Journal: Biotechnology Reports

Article Title: Xenogeneic-free platform for the isolation and scalable expansion of human bladder smooth muscle cells

doi: 10.1016/j.btre.2025.e00878

Figure Lengend Snippet: Isolation and characterization of bladder smooth muscle cells under xeno(geneic)-free culture conditions. A) Samples were fragmented into explants of approximately 1 mm diameter, and 10 explants per well were plated onto 6-well plates (Corning®). Phase contrast imaging of cells migrating and proliferating from explants (first image: donor 63 yo at day 2 post-processing; second image: donor 58 at day 6 post-processing; third and fourth images: donor 70 yo at day 6 post-processing); Scale bar: 100 µm. B) Representative images of fluorescence microscopy for the detection of typical SMC markers (red); left panel: donor 58 yo, cell passage 2; center panel: donor 80 yo, cell passage 5; right panel: donor 70 yo, Passage 3; Nuclei counterstained with DAPI (blue); Scale bar: 100 µm. C) Flow cytometry panel for SMC identity markers (positive and negative) for all four donors. αSMA: α-Smooth muscle actin; MYH11: Myosin heavy chain 11; SM22: Smooth Muscle Protein 22-α; SMC: Smooth muscle cells.

Article Snippet: Primary antibodies anti-human α-Smooth muscle actin (αSMA) (Invitrogen), Caldesmon, Desmin (Santa Cruz Biotechnology), Smooth Muscle Protein 22-α (SM22α) (Novus Biological), Myosin heavy chain 11 or smooth muscle myosin heavy chain (MYH11) (Abcam), CD44, CD90 and CD29 (Becton Dickinson, BD), were diluted in 1% (v/v) BSA and incubated for 2 h at RT.

Techniques: Isolation, Imaging, Fluorescence, Microscopy, Flow Cytometry

Journal: Biotechnology Reports

Article Title: Xenogeneic-free platform for the isolation and scalable expansion of human bladder smooth muscle cells

doi: 10.1016/j.btre.2025.e00878

Figure Lengend Snippet: Assessment of smooth muscle cell identity post-spinner flask culture. A) Live/dead staining assessed by flow cytometry immediately upon cell retrieval at the end of the spinner flask cultures for each donor. B) Flow cytometry for the depicted cell identity markers (positive and negative), C) Representative immunofluorescence images for SMC markers (red) and nuclei counterstained with DAPI (blue) on cells retrieved from microcarriers and adhered to static cultures; Upper panel: donor 58 yo; Middle panel: donor 80 yo; Bottom panel: donor 58 yo (for CD29 and CD44) and donor 80 yo (for CD90); Scale bar: 100 μm. αSMA: α-Smooth muscle actin; MYH11: Myosin heavy chain 11; SM22: Smooth Muscle Protein 22-α, SMC: smooth muscle cells.

Article Snippet: Primary antibodies anti-human α-Smooth muscle actin (αSMA) (Invitrogen), Caldesmon, Desmin (Santa Cruz Biotechnology), Smooth Muscle Protein 22-α (SM22α) (Novus Biological), Myosin heavy chain 11 or smooth muscle myosin heavy chain (MYH11) (Abcam), CD44, CD90 and CD29 (Becton Dickinson, BD), were diluted in 1% (v/v) BSA and incubated for 2 h at RT.

Techniques: Staining, Flow Cytometry, Cell Retrieval, Immunofluorescence

Journal: Advanced Materials Interfaces

Article Title: Preparation of ROS‐Responsive Exosome Coating of Nitinol Material for Making the Neurointerventional Stent

doi: 10.1002/admi.202300028

Figure Lengend Snippet: Figure 6. A) Expression of contractile VSMC marker proteins α-SMA and SM22α on the surface of BN and SMEN samples (scale bar = 50 µm). B) Fluorescence quantification of marker proteins α-SMA and SM22α. C) Transwell VSMCs–HUVECs co-culture effects of two groups of samples on VSMC migration (scale bar = 2 mm). *P ≤ 0.05 and **P ≤ 0.01.

Article Snippet: The cell polarization of VSMCs was induced with a serum-free medium containing platelet-derived growth factor BB subunits (HEGFP-16011; Oricell, Guangzhou, China) at 10 ng mL−1 for 24 h.[21] Then, the cell samples were washed, fixed, and incubated overnight with the primary antibody Smooth Muscle Protein 22-α (SM22α) (10493-1-AP, Proteintech, RRID: AB 2 199 363) or alpha smooth muscle actin (α-SMA) (14395-1-AP, Proteintech, RRID: AB 2 223 009).

Techniques: Expressing, Marker, Fluorescence, Co-Culture Assay, Migration

Journal: Open Medicine

Article Title: LncRNA XIST regulates atherosclerosis progression in ox-LDL-induced HUVECs

doi: 10.1515/med-2021-0200

Figure Lengend Snippet: Ox-LDL suppressed proliferation and enhanced apoptosis and inflammatory response in HUVECs. (a and b) The cell viability of HUVECs exposed to ox-LDL was assessed by MTT assay. (c) The flow cytometry assay was performed for examining the apoptosis rate of HUVECs treated with 40 μg/mL of ox-LDL for 48 h. (d) The protein expression levels of Bax and Bcl-2 were measured by western blot assay. (e) LDH release in the medium was examined by a commercial kit. (f) The levels of IL-6, IL-1β, and TNF-α were evaluated in the medium by matched kits. (g and h) The qPCR and western blot assays were used to show the expression levels of α-SMA and SM22-α in HUVECs treated with ox-LDL. Data shown are mean ± SD and from three independent experiments. * P < 0.05. Abbreviations: oxidized low-density lipoprotein (ox-LDL), anti-BCL2-Associated X (Bax), anti-B-cell lymphoma-2 (Bcl-2), interleukin 6 (IL-6), interleukin 1β (IL-1β), and tumor necrosis factor α (TNF-α), anti-α-smooth muscle actin (α-SMA), anti-smooth muscle protein 22-α (SM22-α).

Article Snippet: The primary antibodies were listed: anti-BCL2-Associated X (Bax; ab32503; 1:1,000 dilution; Abcam), anti-B-cell lymphoma-2 (Bcl-2; ab32124; 1:1,000 dilution; Abcam), anti-α-smooth muscle actin (α-SMA; ab5694; 1:1,000 dilution; Abcam), anti-smooth muscle protein 22-α (SM22-α; ab14106; 1:1,000 dilution; Abcam), anti-PAPPA (ab174314; 1:1,000 dilution; Abcam), and β-actin (ab179467; 1:3,000 dilution; Abcam).

Techniques: MTT Assay, Flow Cytometry, Expressing, Western Blot

Journal: Open Medicine

Article Title: LncRNA XIST regulates atherosclerosis progression in ox-LDL-induced HUVECs

doi: 10.1515/med-2021-0200

Figure Lengend Snippet: Knockdown of XIST weakened ox-LDL-induced effects on HUVECs. (a) The expression level of XIST in HUVECs treated with ox-LDL was determined by qPCR analysis. (b) The interference efficiency of si-XIST in HUVECs was assessed by qPCR assay. (c–f) HUVECs were divided into two groups: ox-LDL + si-NC and ox-LDL + si-XIST groups. (c) The proliferation capability of HUVECs was measured using MTT assay. (d) The flow cytometry assay was recruited to monitor the apoptosis of HUVECs. (e) The western blot analysis was applied to assess Bax and Bcl-2 expression in HUVECs. (f) A commercial available kit was used to assess LDH release in HUVECs supernatants. (g) The expression levels of IL-6, IL-1β, and TNF-α in ox-LDL-induced HUVECs were displayed, with 0 μg/mL of ox-LDL group as control. (h and i) QPCR and western blot analyses were employed to quantify mRNA and protein levels of α-SMA and SM22-α in HUVECs, respectively. Data shown are mean ± SD and from three independent experiments. * P < 0.05. Abbreviations: X-inactive-specific transcript (XIST).

Article Snippet: The primary antibodies were listed: anti-BCL2-Associated X (Bax; ab32503; 1:1,000 dilution; Abcam), anti-B-cell lymphoma-2 (Bcl-2; ab32124; 1:1,000 dilution; Abcam), anti-α-smooth muscle actin (α-SMA; ab5694; 1:1,000 dilution; Abcam), anti-smooth muscle protein 22-α (SM22-α; ab14106; 1:1,000 dilution; Abcam), anti-PAPPA (ab174314; 1:1,000 dilution; Abcam), and β-actin (ab179467; 1:3,000 dilution; Abcam).

Techniques: Expressing, MTT Assay, Flow Cytometry, Western Blot

Journal: Open Medicine

Article Title: LncRNA XIST regulates atherosclerosis progression in ox-LDL-induced HUVECs

doi: 10.1515/med-2021-0200

Figure Lengend Snippet: Ox-LDL repressed proliferation and induced apoptosis and inflammatory response in HUVECs by targeting the XIST/miR-98-5p axis. (a) QPCR was enforced to confirm the overexpression efficiency of miR-98-5p in HUVECs. (b–i) HUVECs were treated with ox-LDL + miR-NC, ox-LDL + miR-98-5p, ox-LDL + miR-98-5p + pcDNA, or ox-LDL + miR-98-5p + pcDNA-XIST. (b) MTT assay was used to assess cell viability of HUVECs. (c) Cells apoptosis assay was performed in transfected HUVECs by flow cytometry assay. (d and e) The protein expression levels of Bax and Bcl-2 were calculated in HUVECs by western blot assay. (f) The LDH release was displayed in the different groups by kit assay. (g) The concentrations of IL-6, IL-1β, and TNF-α in the medium were presented by ELISA assay. (h and i) The mRNA and protein expression levels of α-SMA and SM22-α in HUVECs were detected by qPCR and western blot assays, respectively. Data shown are mean ± SD and from three independent experiments. * P < 0.05.

Article Snippet: The primary antibodies were listed: anti-BCL2-Associated X (Bax; ab32503; 1:1,000 dilution; Abcam), anti-B-cell lymphoma-2 (Bcl-2; ab32124; 1:1,000 dilution; Abcam), anti-α-smooth muscle actin (α-SMA; ab5694; 1:1,000 dilution; Abcam), anti-smooth muscle protein 22-α (SM22-α; ab14106; 1:1,000 dilution; Abcam), anti-PAPPA (ab174314; 1:1,000 dilution; Abcam), and β-actin (ab179467; 1:3,000 dilution; Abcam).

Techniques: Over Expression, MTT Assay, Apoptosis Assay, Transfection, Flow Cytometry, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Open Medicine

Article Title: LncRNA XIST regulates atherosclerosis progression in ox-LDL-induced HUVECs

doi: 10.1515/med-2021-0200

Figure Lengend Snippet: Knockdown of miR-98-5p reversed si-PAPPA induced the effect on proliferation, apoptosis, and inflammatory response in HUVECs. (a and b) The interference efficiency of si-PAPPA in HUVECs was measured using qPCR and western blot assays. (c–f) HUVECs were treated with ox-LDL + si-NC, ox-LDL + si-PAPPA, ox-LDL + si-PAPPA + anti-NC, or ox-LDL + si-PAPPA + anti-miR-98-5p. (c) The cell viability of HUVECs was estimated by MTT assay. (d) Apoptosis analysis was performed in HUVECs using flow cytometry assay. (e) The western blot assay was carried out to analyze Bax and Bcl-2 expression in HUVECs. (f) The LDH detection kit was used to test LDH release in HUVECs. (g) ELISA assay was applied to measure the levels of IL-6, IL-1β, and TNF-α in the medium, with 0 μg/mL of ox-LDL group as control. (h and i) The expression levels of α-SMA and SM22-α in HUVECs were assessed by qPCR and western blot assays. Data shown are mean ± SD and from three independent experiments. * P < 0.05.

Article Snippet: The primary antibodies were listed: anti-BCL2-Associated X (Bax; ab32503; 1:1,000 dilution; Abcam), anti-B-cell lymphoma-2 (Bcl-2; ab32124; 1:1,000 dilution; Abcam), anti-α-smooth muscle actin (α-SMA; ab5694; 1:1,000 dilution; Abcam), anti-smooth muscle protein 22-α (SM22-α; ab14106; 1:1,000 dilution; Abcam), anti-PAPPA (ab174314; 1:1,000 dilution; Abcam), and β-actin (ab179467; 1:3,000 dilution; Abcam).

Techniques: Western Blot, MTT Assay, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay

Journal: British Journal of Pharmacology

Article Title: Inhibition of polycomb repressor complex 2 ameliorates neointimal hyperplasia by suppressing trimethylation of H3K27 in vascular smooth muscle cells

doi: 10.1111/bph.14754

Figure Lengend Snippet: Trimethylation of H3K27 was induced in PDGF‐BB treated vascular smooth muscle cells (VSMCs) and neointima of rat wire‐injured carotid arteries. (a) VSMCs were treated with PDGF‐BB (40 ng·ml−1) for 36 hr after serum starvation. Histone methylation was detected by NANO‐HPLC/MS. The change in methylation is shown as a heat map. (b,c) VSMCs were treated with 10% FBS (b) or PDGF‐BB (40 ng·ml−1) (c) for 36 hr after being starved for 48 hr: western blot analysis of protein levels of H3K27me3, total H3, and GAPDH; data are mean ± SEM, n = 5. (d,e) Rats underwent carotid artery wire injury (n = 5 rats in each group). At 14 days after surgery, arteries were collected. (d) Representative H&E staining and immunostaining of H3K27me3, α‐SMA, and DAPI (left panel, scale bar = 50 μm); quantification of mean integrated OD of H3K27me3 in α‐SMA positive area (right panel). (e) Western blot analysis of protein levels of H3K27me3, total H3, α‐SMA, SM‐22, PCNA, and α‐tubulin. Data are mean ± SEM. *P < .05. Ctrl, control

Article Snippet: Proteins from VSMCs or rat carotid arteries were resolved by 10% or 12% SDS‐PAGE and transferred to PVDF membranes, which were blocked with 5% non‐fat milk for 2 hr at room temperature, then incubated with primary antibodies for H3K27me3 (1:5,000, cat#: 07449), EZH1 (1:2,500, cat#: ABE281), and EZH2 (1:1,000, cat#: 07689, RRID:AB_417397; all from Millipore); total H3 (1:3,000, cat#: 4499, RRID:AB_10544537) and PCNA (1:1,000, cat#: 2586, RRID:AB_2160343; both from CST); α‐SMA (1:8,000, cat#: A2547; Sigma Aldrich); smooth muscle protein 22‐α (SM‐22α; 1:500, cat#: SC‐53932, RRID:AB_1129519) and α‐tubulin (1:1,000, cat#: SC‐8035, RRID:AB_628408; both from Santa Cruz Biotechnology, CA, USA); and GAPDH (1:10,000, cat#: 600041, RRID:AB_2107436; Proteintech) at 4°C overnight, then secondary antibodies (goat anti‐mouse IgG, 1:5,000, cat#: 4741806, RRID:AB_2307348; goat anti‐rabbit IgG, 1:5,000, cat#: 0741506, RRID:AB_2721169; KPL, Gaithersburg, MD, USA) were applied at room temperature for 1 hr.

Techniques: Methylation, Western Blot, Staining, Immunostaining, Control