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myh11 (Proteintech)

Name: myh11
Brand: Proteintech
Rating: 94 (97 reviews)

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Proteintech myh11
Myh11, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/myh11/product/Proteintech
Average 94 stars, based on 97 article reviews

myh11 - by Bioz Stars, 2026-03

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Journal: bioRxiv

Article Title: Ionic Regulation of Mechanosurveillance and Metastasis via the MRTFA/KCNMB1 Axis

doi: 10.64898/2026.01.13.699089

Figure Lengend Snippet: (A) Diagram depicting different MRTFA partners that can promote cortical stiffness in cancer cells. (B) Brightfield images of E0771 cells overexpressing MRTFA WT treated with DMSO (vehicle only control), TGFβ inhibitor (SB-505124; 2.5 µM) or YAP inhibitor (Verteporfin; 10 µM) for 24 hrs. Brightfield images of E0771 cells overexpressing MRTFA WT in the absence or presence of shRNA mediated SRF knockdown (shSRF). Scale bars: 50 µm. (C) Relative transcript levels of downstream MRTFA target genes, Acta2 and Myh11 . Data are shown as fold change calculated from 3 technical replicates per sample and normalized GAPDH. Each graph is representative of 3 independent experiments. (D) Confocal images of representative cells stained with phalloidin (F-actin). Scale bar = 10 µm. (E) Anisotropy measurements for cell lines in (D). Each graph is a compilation of 3 independent experiments; AT-3: pRetroX = 95 cells, MRTFA WT = 95 cells, MRTFA Y238A = 92 cells; E0771: Control (pRetroX) = 165 cells, MRTFA WT =112 cells, MRTFA Y238A =158 cells; Kruskal-Wallace with multiple comparisons. All error bars are mean±s.e.m. ns, not significant; * P <0.05; ** P <0.01; *** P <0.001. (F) Raw AFM stiffness measurements comparing MRTFA WT and MRTFA Y238A compared to control cells. Each graph is a compilation of 2 independent experiments. AT-3: pRetroX = 19 cells, MRTFA WT = 19 cells, MRTFA Y238A = 19 cells; E0771: Control (pRetroX) = 15 cells, MRTFA WT = 16 cells, MRTFA Y238A = 15 cells; Kruskal-Wallace with multiple comparisons. All error bars are mean±s.e.m. ns, not significant; * P <0.05; ** P <0.01; *** P <0.001. (G) Raw AFM stiffness measurements comparing SRF knockdown effect on MRTFA WT cell stiffness. Results are combined from 2 independent experiments. E0771: Control (pRetroX) = 10 cells, MRTFA WT + Control (pLKO) = 11 cells, MRTFA WT + shSRF=10 cells; Kruskal-Wallace with multiple comparisons. All error bars are mean±s.e.m. ns, not significant; * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: TaqMan probes used were all purchased from Thermo Scientific: Gapdh (Mm99999915_g1), Acta2 (Mm00725412_s1), Actg2 (Mm00656102_m1), Tagln (Mm00441661_g1), Myh11 (Mm00443013_m1) , Mrtfa (Mm00461840_m1), Mrtfb (Mm00463877_m1), Srf (Mm00491032_m1), Kcnmb1 (Mm00466621_m1), Ccn2 (Mm01192933_g1), GAPDH (Hs99999905_m1), MRTFA (Hs01090249_g1), MRTFB (Hs00401867_m1), SRF (Hs00182371_m1), KCNMB1 (Hs00188073_m1).

Techniques: Control, shRNA, Knockdown, Staining

(A) Immunoblot of YAP in E0771 MRTFA WT cells treated with 10 µM verteporfin for 24 hours compared to DMSO control. GAPDH is used as loading control. (B) Immunoblot of SMAD phosphorylation in E0771 MRTFA WT cells treated with 2.5ng/mL TGFß2 for 1 hour in the presence or absence of 2.5 µM SB-505124 pre-treatment. GAPDH is used as loading control. (C) MRTFA and MRTFB amino acid alignment around the conserved Y238 and Y305 residues, respectively. (D) Immunoblot of MRTFA overexpression in MRTFA WT and MRTFA Y238A cells compared to control. GAPDH is used as loading control. * indicates endogenous MRTFA. (E) Relative transcript levels of actin-related established MRTFA target genes Actg2 and Myh11 in MRTFA WT and MRTFA Y238A mutant overexpressing cell lines. Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH. Each graph is representative of 3 independent experiments. (F) Representative immunofluorescent images depicting MRTFA protein localization in MRTFA WT and MRTFA Y238A mutant overexpressing cell lines. Scale bars = 50 µm. (G) Quantitation of MRTFA protein localization in MRTFA WT and mutant overexpressing cell lines. AT-3: Control (pRetroX) = 298 cells, MRTFA WT = 140 cells, MRTFA Y238A = 169 cells; E0771: Control (pRetroX) = 185 cells, MRTFA WT = 315 cells, MRTFA Y238A = 232 cells. (H) Relative transcript levels of SRF in E0771 MRTFA WT cell lines with/without SRF knockdown (shSRF). Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH. Graph is representative of 3 independent experiments.

Journal: bioRxiv

Article Title: Ionic Regulation of Mechanosurveillance and Metastasis via the MRTFA/KCNMB1 Axis

doi: 10.64898/2026.01.13.699089

Figure Lengend Snippet: (A) Immunoblot of YAP in E0771 MRTFA WT cells treated with 10 µM verteporfin for 24 hours compared to DMSO control. GAPDH is used as loading control. (B) Immunoblot of SMAD phosphorylation in E0771 MRTFA WT cells treated with 2.5ng/mL TGFß2 for 1 hour in the presence or absence of 2.5 µM SB-505124 pre-treatment. GAPDH is used as loading control. (C) MRTFA and MRTFB amino acid alignment around the conserved Y238 and Y305 residues, respectively. (D) Immunoblot of MRTFA overexpression in MRTFA WT and MRTFA Y238A cells compared to control. GAPDH is used as loading control. * indicates endogenous MRTFA. (E) Relative transcript levels of actin-related established MRTFA target genes Actg2 and Myh11 in MRTFA WT and MRTFA Y238A mutant overexpressing cell lines. Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH. Each graph is representative of 3 independent experiments. (F) Representative immunofluorescent images depicting MRTFA protein localization in MRTFA WT and MRTFA Y238A mutant overexpressing cell lines. Scale bars = 50 µm. (G) Quantitation of MRTFA protein localization in MRTFA WT and mutant overexpressing cell lines. AT-3: Control (pRetroX) = 298 cells, MRTFA WT = 140 cells, MRTFA Y238A = 169 cells; E0771: Control (pRetroX) = 185 cells, MRTFA WT = 315 cells, MRTFA Y238A = 232 cells. (H) Relative transcript levels of SRF in E0771 MRTFA WT cell lines with/without SRF knockdown (shSRF). Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH. Graph is representative of 3 independent experiments.

Techniques: Western Blot, Control, Phospho-proteomics, Over Expression, Mutagenesis, Quantitation Assay, Knockdown

(A, B) Immunofluorescent staining and mean fluorescence intensity (MFI) of SIRT1 (red) in heart (A) and abdominal aorta (B) tissues of PBS and LCWE-injected mice at 2 weeks post-LCWE injection (n=4 to 5/group). DAPI (Blue) used to stain nuclei. Scale bars, 50μm. (C) mRNA levels of Sirt1 measured by qRT-PCR in heart and abdominal aorta tissues of PBS and LCWE-injected mice at 2 weeks post-LCWE injection (n=9/group). Data are presented as mean ± SEM, representative of one experiment (A, B) or pooled from 2 independent experiments (C). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by unpaired t-test (B, C, D, E).

Journal: bioRxiv

Article Title: Sirtuin 1 Activation Mitigates Murine Vasculitis Severity by Promoting Autophagy and Mitophagy

doi: 10.1101/2025.09.04.671113

Figure Lengend Snippet: (A, B) Immunofluorescent staining and mean fluorescence intensity (MFI) of SIRT1 (red) in heart (A) and abdominal aorta (B) tissues of PBS and LCWE-injected mice at 2 weeks post-LCWE injection (n=4 to 5/group). DAPI (Blue) used to stain nuclei. Scale bars, 50μm. (C) mRNA levels of Sirt1 measured by qRT-PCR in heart and abdominal aorta tissues of PBS and LCWE-injected mice at 2 weeks post-LCWE injection (n=9/group). Data are presented as mean ± SEM, representative of one experiment (A, B) or pooled from 2 independent experiments (C). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by unpaired t-test (B, C, D, E).

Article Snippet: To induce SIRT1 deletion, primary smooth muscle cells isolated from Sirt1 fl/fl and Myh11 Cre/ERT2 Sirt1 Δ/Δ mice were treated with 1μM 4-Hydroxytamoxifen (4-OHT; MedChemExpress HY-16950) for 48 hours.

Techniques: Staining, Fluorescence, Injection, Quantitative RT-PCR

(A) Representative H&E-stained heart sections and heart vessel inflammation scores of PBS or LCWE-injected WT and Sirt1 super mice, at 2 weeks post-injection. (n=5 to 10/group). Scale bars, 500μm. (B, C) Representative pictures of the abdominal aorta areas (B), maximal abdominal aorta diameter, and abdominal aorta area measurements (C) of PBS or LCWE-injected WT and Sirt1 super mice, at 2 weeks post-injection. (n=5 to 10/group). (D) IL-1β measurements in the peritoneal lavage of WT and Sirt1 super mice injected with PBS or LCWE 24 hours post-injection. (E) Schematic of the experimental design. (F-H) WT and Sirt1 ER/ER mice were injected with either PBS or LCWE, and one week later, injected with tamoxifen for 5 consecutive days. Heart and abdominal aorta tissues were collected at day 18 post-LCWE injection for analysis. (F) Representative H&E-stained heart sections and heart vessel inflammation scores of PBS or LCWE-injected, tamoxifen-treated WT and Sirt1 super mice, at day 18 post-injection. (n=5 to 14/group). Scale bars, 500μm. (G, H) Representative pictures of the abdominal aorta areas (G), maximal abdominal aorta diameter and abdominal aorta area measurements (H) of PBS or LCWE-injected, tamoxifen-treated WT and Sirt1 ER/ER mice, at day 18 post-injection (n=5 to 14/group). Data are presented as mean ± SEM, representative of at least 2 independent experiments (A-I). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by two-way ANOVA with Tukey post hoc test (A, C, D, F, H).

Journal: bioRxiv

Article Title: Sirtuin 1 Activation Mitigates Murine Vasculitis Severity by Promoting Autophagy and Mitophagy

doi: 10.1101/2025.09.04.671113

Figure Lengend Snippet: (A) Representative H&E-stained heart sections and heart vessel inflammation scores of PBS or LCWE-injected WT and Sirt1 super mice, at 2 weeks post-injection. (n=5 to 10/group). Scale bars, 500μm. (B, C) Representative pictures of the abdominal aorta areas (B), maximal abdominal aorta diameter, and abdominal aorta area measurements (C) of PBS or LCWE-injected WT and Sirt1 super mice, at 2 weeks post-injection. (n=5 to 10/group). (D) IL-1β measurements in the peritoneal lavage of WT and Sirt1 super mice injected with PBS or LCWE 24 hours post-injection. (E) Schematic of the experimental design. (F-H) WT and Sirt1 ER/ER mice were injected with either PBS or LCWE, and one week later, injected with tamoxifen for 5 consecutive days. Heart and abdominal aorta tissues were collected at day 18 post-LCWE injection for analysis. (F) Representative H&E-stained heart sections and heart vessel inflammation scores of PBS or LCWE-injected, tamoxifen-treated WT and Sirt1 super mice, at day 18 post-injection. (n=5 to 14/group). Scale bars, 500μm. (G, H) Representative pictures of the abdominal aorta areas (G), maximal abdominal aorta diameter and abdominal aorta area measurements (H) of PBS or LCWE-injected, tamoxifen-treated WT and Sirt1 ER/ER mice, at day 18 post-injection (n=5 to 14/group). Data are presented as mean ± SEM, representative of at least 2 independent experiments (A-I). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by two-way ANOVA with Tukey post hoc test (A, C, D, F, H).

Techniques: Staining, Injection

(A) Principal component (PC) analysis of the proteome from the abdominal aortas of WT and Sirt1 super mice injected with either PBS or LCWE, at 2 weeks post-injection (n=5/group). (B) Venn diagram of differentially expressed proteins (DEPs, p-value <0.05; fold change [FC] >2) between the indicated groups. A set of 443 proteins associated with the development of LCWE-induced KD was identified (highlighted in red). (C) Selected pathways and functional annotation terms of proteins increased in LCWE-injected WT mice compared with PBS-injected WT mice and decreased in LCWE-injected Sirt1 super mice compared with LCWE-injected WT mice (n=5/group). (D) Selected pathways and functional annotation terms of proteins decreased in LCWE-injected WT mice compared with PBS-injected WT mice and increased in LCWE-injected Sirt1 super mice compared with LCWE-injected WT mice (n=5/group). (E) Ingenuity pathway analysis (IPA) performed on the set of DEPs associated with LCWE-induced KD highlighted in red from (B) between the indicated groups. (F) Mitochondrial respiration was analyzed in isolated mitochondria from heart tissues of LCWE-injected WT and Sirt1 super mice. Measurements of acute response (left), coupling efficiency (middle), and ATP production (right). Data are presented as mean ± SEM, representative of one experiment. Each symbol represents one individual mouse. * p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by unpaired t-test (F).

Journal: bioRxiv

Article Title: Sirtuin 1 Activation Mitigates Murine Vasculitis Severity by Promoting Autophagy and Mitophagy

doi: 10.1101/2025.09.04.671113

Figure Lengend Snippet: (A) Principal component (PC) analysis of the proteome from the abdominal aortas of WT and Sirt1 super mice injected with either PBS or LCWE, at 2 weeks post-injection (n=5/group). (B) Venn diagram of differentially expressed proteins (DEPs, p-value <0.05; fold change [FC] >2) between the indicated groups. A set of 443 proteins associated with the development of LCWE-induced KD was identified (highlighted in red). (C) Selected pathways and functional annotation terms of proteins increased in LCWE-injected WT mice compared with PBS-injected WT mice and decreased in LCWE-injected Sirt1 super mice compared with LCWE-injected WT mice (n=5/group). (D) Selected pathways and functional annotation terms of proteins decreased in LCWE-injected WT mice compared with PBS-injected WT mice and increased in LCWE-injected Sirt1 super mice compared with LCWE-injected WT mice (n=5/group). (E) Ingenuity pathway analysis (IPA) performed on the set of DEPs associated with LCWE-induced KD highlighted in red from (B) between the indicated groups. (F) Mitochondrial respiration was analyzed in isolated mitochondria from heart tissues of LCWE-injected WT and Sirt1 super mice. Measurements of acute response (left), coupling efficiency (middle), and ATP production (right). Data are presented as mean ± SEM, representative of one experiment. Each symbol represents one individual mouse. * p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by unpaired t-test (F).

Techniques: Injection, Functional Assay, Isolation

(A) Quantification of dihydroethidium (DHE) staining of heart (top) and abdominal aorta (bottom) tissues from WT and Sirt1 super mice injected with either PBS or LCWE at 2 weeks post-injection (n=4 to 5/group). (B) Heatmap of differentially expressed proteins related to autophagy/ROS, fatty acid oxidation, and oxidative phosphorylation pathways between abdominal aortas of PBS and LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=5/group). (C) Immunofluorescent staining and quantification of p62 (red) and LC3B puncta (red) in abdominal aorta tissues from PBS or LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=9,10/group). DAPI (blue) is used to stain nuclei. Scale bars, 50μm. (D) Immunofluorescent staining and quantification of p62 (red) and LC3B puncta (green) from hearts of PBS or LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=9,10/group). DAPI (blue) is used to stain nuclei. Scale bars, 50μm. (E) Western blot analysis of SIRT1, pUbiquitin S65 and β-actin in protein lysates from heart tissues from WT and Sirt1 super mice injected with LCWE at 2 weeks post-injection (n=4, 5/group). Data are presented as mean ± SEM. Pooled from 2 experiments (A, C, D, E), representative of one experiment (B). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by two-way ANOVA with Tukey post hoc test (A) or unpaired t-test (C, D). MFI, mean fluorescence intensity.

Journal: bioRxiv

Article Title: Sirtuin 1 Activation Mitigates Murine Vasculitis Severity by Promoting Autophagy and Mitophagy

doi: 10.1101/2025.09.04.671113

Figure Lengend Snippet: (A) Quantification of dihydroethidium (DHE) staining of heart (top) and abdominal aorta (bottom) tissues from WT and Sirt1 super mice injected with either PBS or LCWE at 2 weeks post-injection (n=4 to 5/group). (B) Heatmap of differentially expressed proteins related to autophagy/ROS, fatty acid oxidation, and oxidative phosphorylation pathways between abdominal aortas of PBS and LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=5/group). (C) Immunofluorescent staining and quantification of p62 (red) and LC3B puncta (red) in abdominal aorta tissues from PBS or LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=9,10/group). DAPI (blue) is used to stain nuclei. Scale bars, 50μm. (D) Immunofluorescent staining and quantification of p62 (red) and LC3B puncta (green) from hearts of PBS or LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=9,10/group). DAPI (blue) is used to stain nuclei. Scale bars, 50μm. (E) Western blot analysis of SIRT1, pUbiquitin S65 and β-actin in protein lysates from heart tissues from WT and Sirt1 super mice injected with LCWE at 2 weeks post-injection (n=4, 5/group). Data are presented as mean ± SEM. Pooled from 2 experiments (A, C, D, E), representative of one experiment (B). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by two-way ANOVA with Tukey post hoc test (A) or unpaired t-test (C, D). MFI, mean fluorescence intensity.

Techniques: Staining, Injection, Phospho-proteomics, Western Blot, Fluorescence

$(A) Heatmap of differentially expressed proteins (DEPs) related to VSMCs type I and synthetic VSMCs type II in the abdominal aortas of PBS and LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=5/group). (B) Primary coronary artery SMCs were treated with SRT1720 (5μM) or EX527 (5μM) alone or in combination with LCWE (60μg/ml, 24 hours). Representative Western blots of LC3-I/II, pS6 S235/236 , S6, pULK1 S555 , pAMPK T172 and β-actin from whole cell lysate (n=4/group). (C) Representative Western blots of p62, LC3-I/II, TOM70 and Rho-GDI from Mitochondrial (Mito) and cytosolic (Cyto) fractions of primary coronary artery SMCs treated with SRT1720 (5μM) or EX527 (5μM) alone or in combination with LCWE (60μg/ml, 24 hours) and CQ (5μM, last 4 hours). Ponceau S. was used as loading control. (D) Representative Western blots of LC3-I/II, TOM70, COXIV and β-actin from whole cell lysate of primary coronary artery SMCs treated with SRT1720 (5μM) or EX527 (5μM) alone or in combination with LCWE (60μg/ml, 24 hours) and CQ (5μM, last 4 hours). (E) Quantification of CytoID staining in WT primary coronary artery SMCs treated with SRT1720 (5μM, 24 hours) or EX527 (5μM, 24 hours) alone or in combination with rIL-1β (10ng/ml, 24 hours) and CQ (5μM, last 4 hours) (n=3/group). (F) Quantification of mitoSOX/mitoTracker ratio from WT Primary coronary artery SMCs treated with SRT1720 (5μM, 24 hours) or EX527 (5μM, 24 hours) alone or in combination with rIL-1β (10ng/ml, 24 hours) (n=8/group). (G-H) Bone marrow-derived macrophages (BMDMs) differentiated from WT and Sirt1 super mice treated with LCWE (60μg/ml, 24 hours). (G) IL-1β and TNFα levels were measured from the supernatants by ELISA (n=6/group). (H) Representative Western blots for SIRT1 and pS6 S235/236 from whole cell lysates (n=4/group). Ponceau S. was used as loading control. Data are presented as mean ± SEM. Representative of 2 experiments (B, C, D, E, F, G, H), representative of one experiment (A). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by one-way ANOVA (E, F) or two-way ANOVA with Tukey post hoc test (H). rIL-1β, recombinant IL-1β.$

Journal: bioRxiv

Article Title: Sirtuin 1 Activation Mitigates Murine Vasculitis Severity by Promoting Autophagy and Mitophagy

doi: 10.1101/2025.09.04.671113

Figure Lengend Snippet: (A) Heatmap of differentially expressed proteins (DEPs) related to VSMCs type I and synthetic VSMCs type II in the abdominal aortas of PBS and LCWE-injected WT and Sirt1 super mice at 2 weeks post-injection (n=5/group). (B) Primary coronary artery SMCs were treated with SRT1720 (5μM) or EX527 (5μM) alone or in combination with LCWE (60μg/ml, 24 hours). Representative Western blots of LC3-I/II, pS6 S235/236 , S6, pULK1 S555 , pAMPK T172 and β-actin from whole cell lysate (n=4/group). (C) Representative Western blots of p62, LC3-I/II, TOM70 and Rho-GDI from Mitochondrial (Mito) and cytosolic (Cyto) fractions of primary coronary artery SMCs treated with SRT1720 (5μM) or EX527 (5μM) alone or in combination with LCWE (60μg/ml, 24 hours) and CQ (5μM, last 4 hours). Ponceau S. was used as loading control. (D) Representative Western blots of LC3-I/II, TOM70, COXIV and β-actin from whole cell lysate of primary coronary artery SMCs treated with SRT1720 (5μM) or EX527 (5μM) alone or in combination with LCWE (60μg/ml, 24 hours) and CQ (5μM, last 4 hours). (E) Quantification of CytoID staining in WT primary coronary artery SMCs treated with SRT1720 (5μM, 24 hours) or EX527 (5μM, 24 hours) alone or in combination with rIL-1β (10ng/ml, 24 hours) and CQ (5μM, last 4 hours) (n=3/group). (F) Quantification of mitoSOX/mitoTracker ratio from WT Primary coronary artery SMCs treated with SRT1720 (5μM, 24 hours) or EX527 (5μM, 24 hours) alone or in combination with rIL-1β (10ng/ml, 24 hours) (n=8/group). (G-H) Bone marrow-derived macrophages (BMDMs) differentiated from WT and Sirt1 super mice treated with LCWE (60μg/ml, 24 hours). (G) IL-1β and TNFα levels were measured from the supernatants by ELISA (n=6/group). (H) Representative Western blots for SIRT1 and pS6 S235/236 from whole cell lysates (n=4/group). Ponceau S. was used as loading control. Data are presented as mean ± SEM. Representative of 2 experiments (B, C, D, E, F, G, H), representative of one experiment (A). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by one-way ANOVA (E, F) or two-way ANOVA with Tukey post hoc test (H). rIL-1β, recombinant IL-1β.

Techniques: Injection, Western Blot, Control, Staining, Derivative Assay, Enzyme-linked Immunosorbent Assay, Recombinant

(A) Representative H&E-stained heart sections and heart vessel inflammation scores of Sirt1 fl/fl and Myh11 Cre/ERT2 Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=9 to 10/group). Scale bars, 500μm. (B, C) Representative pictures of the abdominal aorta areas (B), maximal abdominal aorta diameter and abdominal aorta area (C) measurements of Sirt1 fl/fl and Myh11 Cre/ERT2 Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=9 to 10/group). (D) Levels of IL-1β in the peritoneal lavage of Sirt1 fl/fl and Myh11 Cre/ERT2 Sirt1 Δ/Δ mice injected with PBS or LCWE 24 hours post-injection. (E-G) Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=5 to 11–13/group). (E) Representative H&E-stained heart sections and heart vessel inflammation scores of Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=5 to 11–12/group). Scale bars, 500μm. (F, G) Representative pictures of the abdominal aorta areas (F), maximal abdominal aorta diameter, and abdominal aorta area measurements (G) of Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=5 to 11–12/group). (H) Levels of IL-1β in the peritoneal lavage of Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice injected with PBS or LCWE 24 hours post-injection (n=5/group). (I) Quantification of FLICA + F4/80 + NLRP3 + cell numbers in heart tissues from PBS or LCWE injected Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice at 2 weeks post-injection (n=5 to 7/group). (J) Quantification of FLICA + F4/80 + NLRP3 + cell numbers in abdominal aortas from PBS or LCWE injected Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice at 2 weeks post-injection (n=4 to 5–8/group). Data are presented as mean ± SEM. Pooled from 2 experiments (A-J). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by two-way ANOVA with Tukey post hoc test (A, C, D, E, G, H, I, J). FLICA, Fluorochrome Labeled Inhibitors of Caspases.

Journal: bioRxiv

Article Title: Sirtuin 1 Activation Mitigates Murine Vasculitis Severity by Promoting Autophagy and Mitophagy

doi: 10.1101/2025.09.04.671113

Figure Lengend Snippet: (A) Representative H&E-stained heart sections and heart vessel inflammation scores of Sirt1 fl/fl and Myh11 Cre/ERT2 Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=9 to 10/group). Scale bars, 500μm. (B, C) Representative pictures of the abdominal aorta areas (B), maximal abdominal aorta diameter and abdominal aorta area (C) measurements of Sirt1 fl/fl and Myh11 Cre/ERT2 Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=9 to 10/group). (D) Levels of IL-1β in the peritoneal lavage of Sirt1 fl/fl and Myh11 Cre/ERT2 Sirt1 Δ/Δ mice injected with PBS or LCWE 24 hours post-injection. (E-G) Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=5 to 11–13/group). (E) Representative H&E-stained heart sections and heart vessel inflammation scores of Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=5 to 11–12/group). Scale bars, 500μm. (F, G) Representative pictures of the abdominal aorta areas (F), maximal abdominal aorta diameter, and abdominal aorta area measurements (G) of Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice were injected with PBS or LCWE and analyzed at 2 weeks post-injection (n=5 to 11–12/group). (H) Levels of IL-1β in the peritoneal lavage of Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice injected with PBS or LCWE 24 hours post-injection (n=5/group). (I) Quantification of FLICA + F4/80 + NLRP3 + cell numbers in heart tissues from PBS or LCWE injected Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice at 2 weeks post-injection (n=5 to 7/group). (J) Quantification of FLICA + F4/80 + NLRP3 + cell numbers in abdominal aortas from PBS or LCWE injected Sirt1 fl/fl and Csf1r Cre Sirt1 Δ/Δ mice at 2 weeks post-injection (n=4 to 5–8/group). Data are presented as mean ± SEM. Pooled from 2 experiments (A-J). Each symbol represents one individual mouse. *p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, and **** p-value < 0.001 by two-way ANOVA with Tukey post hoc test (A, C, D, E, G, H, I, J). FLICA, Fluorochrome Labeled Inhibitors of Caspases.

Techniques: Staining, Injection, Labeling

AGE Effects on Human VSMCs: Promotion of Migration and Inhibition of Contraction. (A) ELISA analysis detected the accumulation of AGEs in lysates of human aortic tissues. (B–D) RT-qPCR (B,C) and Western blot (D) detected the expression of the contraction marker α-SMA and the migration marker MMP-2 in human VSMCs after AGE treatment for 24 h; (E) Transwell migration experiment detected the migration of human VSMCs after AGE treatment for 24 h or 72 h; (F) Immunofluorescence detected the expression of the contraction marker MYH11 in human VSMCs after AGE treatment for 24 h. Blue fluorescence indicates DAPI staining, and green indicates MYH11 staining. (G) The CTG assay detected the growth curve of human VSMCs under the treatment with 200 μg/mL AGE. Data are presented as the mean ± SD from three independent experiments and were analyzed with a one-way ANOVA followed by a Dunnett’s multiple comparisons test. Asterisks indicate significant differences (*P < 0.05, **P < 0.01, ***P < 0.001) between the treatment group and the control group.

Journal: Experimental Biology and Medicine

Article Title: Mechanisms of AGE-induced VSMC phenotypic switching and macrophage modulation in human abdominal aortic aneurysms

doi: 10.3389/ebm.2025.10527

Figure Lengend Snippet: AGE Effects on Human VSMCs: Promotion of Migration and Inhibition of Contraction. (A) ELISA analysis detected the accumulation of AGEs in lysates of human aortic tissues. (B–D) RT-qPCR (B,C) and Western blot (D) detected the expression of the contraction marker α-SMA and the migration marker MMP-2 in human VSMCs after AGE treatment for 24 h; (E) Transwell migration experiment detected the migration of human VSMCs after AGE treatment for 24 h or 72 h; (F) Immunofluorescence detected the expression of the contraction marker MYH11 in human VSMCs after AGE treatment for 24 h. Blue fluorescence indicates DAPI staining, and green indicates MYH11 staining. (G) The CTG assay detected the growth curve of human VSMCs under the treatment with 200 μg/mL AGE. Data are presented as the mean ± SD from three independent experiments and were analyzed with a one-way ANOVA followed by a Dunnett’s multiple comparisons test. Asterisks indicate significant differences (*P < 0.05, **P < 0.01, ***P < 0.001) between the treatment group and the control group.

Article Snippet: After blocking with 3% BSA at room temperature for 30 min, the cells were incubated overnight at 4°C with MYH11 Antibody (1:100, Proteintech, Cat. No. 21404-1-AP).

Techniques: Migration, Inhibition, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Expressing, Marker, Immunofluorescence, Fluorescence, Staining, CTG Assay, Control

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