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ATCC mouse mesangial cell line mes13 culture mouse mesangial cell line
Mouse Mesangial Cell Line Mes13 Culture Mouse Mesangial Cell Line, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 128 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse mesangial cell line mes13 culture mouse mesangial cell line - by Bioz Stars, 2026-06

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Figure 1. Involvement of RIPK1/RIPK3/MLKL-dependent necroptosis in the mesangial cells of MRL/lpr. The frozen section of the kidney from 18-week-old MRL/lpr and C57BL/6 mice were used for immunofluorescent staining. (A) The necroptosis happened mainly in podocytes and mesangial cells, but not in endothelial cells. Immunofluorescent colocalization of podocyte (podoplanin), endothelial cells (CD31), and mesangial cells (PDGFR-β) with p-RIPK3 or p-MLKL in the glomerulus of 18-week-old MRL/lpr female mice. (B-D) The expression of p-RIPK1 (B), p-RIPK3 (C), and p-MLKL (D) in the mesangial cells in MRL/lpr and C57BL/6 mice. (E-G) The proportion of positive p-RIPK1 (E), p-RIPK3 (F), or p-MLKL (G) nucleus in each glomerulus in MRL/lpr and C57BL/6 mice. Mesangial cells were labeled with PDGFR-β in green. Podocytes were labeled with podoplanin in green, and endothelial cells were labeled with CD31 in green. p-RIPK1, p-RIPK3, and p-MLKL were stained in red. Scale bar: 20μm. Data in Figures E, F, and G was calculated with Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001.

Journal: Autoimmunity

Article Title: Receptor interacting protein kinase 1 activation and triggering mesangial cells necroptosis in MRL/lpr mice model of lupus nephritis.

doi: 10.1080/08916934.2025.2515825

Figure Lengend Snippet: Figure 1. Involvement of RIPK1/RIPK3/MLKL-dependent necroptosis in the mesangial cells of MRL/lpr. The frozen section of the kidney from 18-week-old MRL/lpr and C57BL/6 mice were used for immunofluorescent staining. (A) The necroptosis happened mainly in podocytes and mesangial cells, but not in endothelial cells. Immunofluorescent colocalization of podocyte (podoplanin), endothelial cells (CD31), and mesangial cells (PDGFR-β) with p-RIPK3 or p-MLKL in the glomerulus of 18-week-old MRL/lpr female mice. (B-D) The expression of p-RIPK1 (B), p-RIPK3 (C), and p-MLKL (D) in the mesangial cells in MRL/lpr and C57BL/6 mice. (E-G) The proportion of positive p-RIPK1 (E), p-RIPK3 (F), or p-MLKL (G) nucleus in each glomerulus in MRL/lpr and C57BL/6 mice. Mesangial cells were labeled with PDGFR-β in green. Podocytes were labeled with podoplanin in green, and endothelial cells were labeled with CD31 in green. p-RIPK1, p-RIPK3, and p-MLKL were stained in red. Scale bar: 20μm. Data in Figures E, F, and G was calculated with Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Cell culture and treatment Mouse mesangial cells (ATCC: SV40 MES13) were cultured in DEME/F12 with 5% fetal bovine serum (Gibco, Cat No.: 26170035) and 1% penicillin-streptomycin at 37 °C in 5% CO2 incubator for growth.

Techniques: Staining, Expressing, Labeling

Figure 2. RIPK1 inhibitor ZJU37 reduced the mesangial cell necroptosis in the glomerulus of MRL/lpr mice. 18-week-old MRL/lpr female mice were subjected to ZJU37 or the corresponding vehicle for 4 weeks. (A) Representative PAS-stained kidney sections indicated glomerular and tubular injury as well as vasculitis in the Vehicle or ZJU37 group, Scale bar: 25um. (B) The histological scores of glomerular injury, tubular injury, and vasculitis were evaluated for each mouse in the Vehicle or ZJU37 group. N = 4 mice. * P < 0.05, **p < 0.01, ***p < 0.001. (C) Representative images of p-RIPK3 or p-MLKL by immunofluorescent staining in Vehicle or ZJU37 group. The mesangial cells were labeled with PDGFR-β (green) and DAPI (blue) was used for nuclear staining, Scale bar = 25um. (D) The ratio of positive p-RIPK3 or p-MLKL nucleus to the total nucleus by immunofluorescent staining in Vehicle and ZJU37 group. 10 glomeruli were chosen for each mouse and we plot the mean of the collected data. N = 4 mice. * P < 0.05, **p < 0.01, ***p < 0.001.

Journal: Autoimmunity

Article Title: Receptor interacting protein kinase 1 activation and triggering mesangial cells necroptosis in MRL/lpr mice model of lupus nephritis.

doi: 10.1080/08916934.2025.2515825

Figure Lengend Snippet: Figure 2. RIPK1 inhibitor ZJU37 reduced the mesangial cell necroptosis in the glomerulus of MRL/lpr mice. 18-week-old MRL/lpr female mice were subjected to ZJU37 or the corresponding vehicle for 4 weeks. (A) Representative PAS-stained kidney sections indicated glomerular and tubular injury as well as vasculitis in the Vehicle or ZJU37 group, Scale bar: 25um. (B) The histological scores of glomerular injury, tubular injury, and vasculitis were evaluated for each mouse in the Vehicle or ZJU37 group. N = 4 mice. * P < 0.05, **p < 0.01, ***p < 0.001. (C) Representative images of p-RIPK3 or p-MLKL by immunofluorescent staining in Vehicle or ZJU37 group. The mesangial cells were labeled with PDGFR-β (green) and DAPI (blue) was used for nuclear staining, Scale bar = 25um. (D) The ratio of positive p-RIPK3 or p-MLKL nucleus to the total nucleus by immunofluorescent staining in Vehicle and ZJU37 group. 10 glomeruli were chosen for each mouse and we plot the mean of the collected data. N = 4 mice. * P < 0.05, **p < 0.01, ***p < 0.001.

Techniques: Staining, Labeling

Figure 3. In Vitro necroptosis and apoptosis of mouse mesangial cell line triggering by TSZ and TNFα. For the necroptosis experiment, mouse mesangial cells were treated with 100 ng/ml TNF-α recombinant protein, 50 μM Z-VAD-FMK, and 50 nM SM-164 (TSZ) to induce necroptosis, and 10 μM ZJU37 to inhibit necroptosis. For the apoptosis experiment, mouse mesangial cells were treated with 100 ng/ml TNF-α recombinant protein to induce apoptosis, and 10 μM ZJU37 to inhibit apoptosis. After culturing for 24 or 48 hours, the cells were sampled for downstream experiments. (A) Cell viability was assessed by Cell Counting-Lite 2.0 assay in the DMSO, ZJU37, TSZ, TSZ + ZJU37, TNFα and TNFα + ZJU37 groups, N = 6. (B) Transmission electron microscopy imaging depicting the vesicles and membranous integrity (by red arrow) in the TSZ and TSZ + ZJU37 groups. (C) Representative immunofluorescent images and positive nuclei proportion of p-RIPK1 and p-MLKL in the DMSO, ZJU37, TSZ, TSZ + ZJU37, TNFα, and TNFα + ZJU37 group. 6 visual fields were counted at random and we plot the mean of the collected data. N = 3. (D, E) Western blotting images and statistical figures of p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL, and MLKL in the DMSO, ZJU37, TSZ, TSZ + ZJU37, TNFα, and TNFα + ZJU37 group, N = 3. Experiments in Figures C and D were repeated twice. Data in Figures A, C, and E was calculated with ANOVA and pairwise comparison. *p < 0.05, **p < 0.01, ***p < 0.001 compared to the vehicle group. ns indicated no significance.

Journal: Autoimmunity

Article Title: Receptor interacting protein kinase 1 activation and triggering mesangial cells necroptosis in MRL/lpr mice model of lupus nephritis.

doi: 10.1080/08916934.2025.2515825

Figure Lengend Snippet: Figure 3. In Vitro necroptosis and apoptosis of mouse mesangial cell line triggering by TSZ and TNFα. For the necroptosis experiment, mouse mesangial cells were treated with 100 ng/ml TNF-α recombinant protein, 50 μM Z-VAD-FMK, and 50 nM SM-164 (TSZ) to induce necroptosis, and 10 μM ZJU37 to inhibit necroptosis. For the apoptosis experiment, mouse mesangial cells were treated with 100 ng/ml TNF-α recombinant protein to induce apoptosis, and 10 μM ZJU37 to inhibit apoptosis. After culturing for 24 or 48 hours, the cells were sampled for downstream experiments. (A) Cell viability was assessed by Cell Counting-Lite 2.0 assay in the DMSO, ZJU37, TSZ, TSZ + ZJU37, TNFα and TNFα + ZJU37 groups, N = 6. (B) Transmission electron microscopy imaging depicting the vesicles and membranous integrity (by red arrow) in the TSZ and TSZ + ZJU37 groups. (C) Representative immunofluorescent images and positive nuclei proportion of p-RIPK1 and p-MLKL in the DMSO, ZJU37, TSZ, TSZ + ZJU37, TNFα, and TNFα + ZJU37 group. 6 visual fields were counted at random and we plot the mean of the collected data. N = 3. (D, E) Western blotting images and statistical figures of p-RIPK1, RIPK1, p-RIPK3, RIPK3, p-MLKL, and MLKL in the DMSO, ZJU37, TSZ, TSZ + ZJU37, TNFα, and TNFα + ZJU37 group, N = 3. Experiments in Figures C and D were repeated twice. Data in Figures A, C, and E was calculated with ANOVA and pairwise comparison. *p < 0.05, **p < 0.01, ***p < 0.001 compared to the vehicle group. ns indicated no significance.

Techniques: In Vitro, Recombinant, Cell Counting, Transmission Assay, Electron Microscopy, Imaging, Western Blot, Comparison

Figure 4. In Vitro necroptosis of mouse mesangial cell line with serum from MRL/lpr mice and RIPK1 inhibitor ZJU37. Mouse mesangial cells were cultured with 100ul serum from 18-week-old MRL/lpr mice or serum + ZJU37 for 24 or 48 hours for subsequent experiments. (A) Cell viability was assessed by Cell Counting-Lite 2.0 assay in the DMSO, serum, and serum + ZJU37 groups. N = 6; (B) Transmission electron microscopy imaging depicting the vesicles and membranous integrity in the DMSO, serum, and serum + ZJU37 groups; (C) Representative immunofluorescent images and positive nuclei proportion of p-RIPK1, p-RIPK3, and p-MLKL in the DMSO, serum, and serum + ZJU37 group. Six visual fields were counted at random and we plot the mean of the collected data. N = 3. (D, E) Western blotting images and statistical figures of p-RIPK1, RIPK1, RIPK3, and p-RIPK3 in the DMSO, serum, and serum + ZJU37 group, N = 3. Experiments in Figures C and D were repeated twice. Data in Figures A, C, and E was calculated with ANOVA and pairwise comparison. *p < 0.05, **p < 0.01, ***p < 0.001 compared to the vehicle group. ns indicated no significance.

Journal: Autoimmunity

Article Title: Receptor interacting protein kinase 1 activation and triggering mesangial cells necroptosis in MRL/lpr mice model of lupus nephritis.

doi: 10.1080/08916934.2025.2515825

Figure Lengend Snippet: Figure 4. In Vitro necroptosis of mouse mesangial cell line with serum from MRL/lpr mice and RIPK1 inhibitor ZJU37. Mouse mesangial cells were cultured with 100ul serum from 18-week-old MRL/lpr mice or serum + ZJU37 for 24 or 48 hours for subsequent experiments. (A) Cell viability was assessed by Cell Counting-Lite 2.0 assay in the DMSO, serum, and serum + ZJU37 groups. N = 6; (B) Transmission electron microscopy imaging depicting the vesicles and membranous integrity in the DMSO, serum, and serum + ZJU37 groups; (C) Representative immunofluorescent images and positive nuclei proportion of p-RIPK1, p-RIPK3, and p-MLKL in the DMSO, serum, and serum + ZJU37 group. Six visual fields were counted at random and we plot the mean of the collected data. N = 3. (D, E) Western blotting images and statistical figures of p-RIPK1, RIPK1, RIPK3, and p-RIPK3 in the DMSO, serum, and serum + ZJU37 group, N = 3. Experiments in Figures C and D were repeated twice. Data in Figures A, C, and E was calculated with ANOVA and pairwise comparison. *p < 0.05, **p < 0.01, ***p < 0.001 compared to the vehicle group. ns indicated no significance.

Techniques: In Vitro, Cell Culture, Cell Counting, Transmission Assay, Electron Microscopy, Imaging, Western Blot, Comparison

Figure 5. The GO and KEGG analysis of the main pathways changed in mesangial cells using serum from MRL/lpr mice and RIPK1 inhibitor ZJU37. Mouse mesangial cell line was stimulated with DMSO, serum from 18-week-old MRL/lpr mice (serum group), or ZJU37 + serum from 18-week-old MRL/ lpr mice (ZJU37 group) for 48 hours. (A) The Venn diagram of the various expression genes in the DMSO, serum, and ZJU37 groups. (B) The heatmap depicts the different expression genes in the DMSO, serum, and ZJU37 groups. The volcano map (C), GO enrichment (E), and top 20 KEGG pathways analysis (G) of up- or down-regulated genes in serum versus DMSO groups. The volcano map (D), GO enrichment (F), and top 20 KEGG pathways analysis (H) of the up-and down-regulated genes in the ZJU37 VS serum group.

Journal: Autoimmunity

Article Title: Receptor interacting protein kinase 1 activation and triggering mesangial cells necroptosis in MRL/lpr mice model of lupus nephritis.

doi: 10.1080/08916934.2025.2515825

Figure Lengend Snippet: Figure 5. The GO and KEGG analysis of the main pathways changed in mesangial cells using serum from MRL/lpr mice and RIPK1 inhibitor ZJU37. Mouse mesangial cell line was stimulated with DMSO, serum from 18-week-old MRL/lpr mice (serum group), or ZJU37 + serum from 18-week-old MRL/ lpr mice (ZJU37 group) for 48 hours. (A) The Venn diagram of the various expression genes in the DMSO, serum, and ZJU37 groups. (B) The heatmap depicts the different expression genes in the DMSO, serum, and ZJU37 groups. The volcano map (C), GO enrichment (E), and top 20 KEGG pathways analysis (G) of up- or down-regulated genes in serum versus DMSO groups. The volcano map (D), GO enrichment (F), and top 20 KEGG pathways analysis (H) of the up-and down-regulated genes in the ZJU37 VS serum group.

Techniques: Expressing

Figure 6. Validation of the function in mouse mesangial cells using serum from MRL/lpr mice, RIPK1 inhibitor ZJU37, PI3K inhib itor LY294002, and anti-mouse IL17A. Mouse mesangial cell line was stimulated with DMSO, serum from 18-week-old MRL/lpr mice (serum group), serum + ZJU37, serum + LY294002, serum + LY294002 + ZJU37, serum + anti-mouse IL17A as well as serum + anti-mouse IL17A + ZJU37 for 24 hours. (A) Representative images of wound healing in 0h, 12h, and 24h in DMSO, serum, and serum + ZJU37 group. (B) The migration rate of mouse mesangial cells in 12h and 24h in DMSO, serum, and serum + ZJU37 group. 6 would areas were counted at random and we plot the mean of the collected data. N = 6. (C, D) Representative immunoflu orescent images and the proportion of positive EdU nucleus in the DMSO, serum, and serum + ZJU37 group. 6 visual fields were counted at random and we plot the mean of the collected data. N = 6. (E, F) Western blotting images and statistical figures of PI3K, p-AKT, and AKT in the DMSO, serum, serum + ZJU37, serum + LY294002, and serum + LY294002 + ZJU37 group. N = 3. (G, H) Western blotting images and statistical figures of p-NFκB, NFκB, p-ERK, and ERK in the DMSO, serum, serum + ZJU37, serum + anti-mouse IL17A, and serum + anti-mouse IL17A + ZJU37 group. N = 3. Experiments in Figures E and G were repeated twice. Data in Figures B, D, F, and H was calculated with ANOVA and pairwise comparison. *p < 0.05, **p < 0.01, ***p < 0.001 com pared to the vehicle group. ns indicated no significance.

Journal: Autoimmunity

Article Title: Receptor interacting protein kinase 1 activation and triggering mesangial cells necroptosis in MRL/lpr mice model of lupus nephritis.

doi: 10.1080/08916934.2025.2515825

Figure Lengend Snippet: Figure 6. Validation of the function in mouse mesangial cells using serum from MRL/lpr mice, RIPK1 inhibitor ZJU37, PI3K inhib itor LY294002, and anti-mouse IL17A. Mouse mesangial cell line was stimulated with DMSO, serum from 18-week-old MRL/lpr mice (serum group), serum + ZJU37, serum + LY294002, serum + LY294002 + ZJU37, serum + anti-mouse IL17A as well as serum + anti-mouse IL17A + ZJU37 for 24 hours. (A) Representative images of wound healing in 0h, 12h, and 24h in DMSO, serum, and serum + ZJU37 group. (B) The migration rate of mouse mesangial cells in 12h and 24h in DMSO, serum, and serum + ZJU37 group. 6 would areas were counted at random and we plot the mean of the collected data. N = 6. (C, D) Representative immunoflu orescent images and the proportion of positive EdU nucleus in the DMSO, serum, and serum + ZJU37 group. 6 visual fields were counted at random and we plot the mean of the collected data. N = 6. (E, F) Western blotting images and statistical figures of PI3K, p-AKT, and AKT in the DMSO, serum, serum + ZJU37, serum + LY294002, and serum + LY294002 + ZJU37 group. N = 3. (G, H) Western blotting images and statistical figures of p-NFκB, NFκB, p-ERK, and ERK in the DMSO, serum, serum + ZJU37, serum + anti-mouse IL17A, and serum + anti-mouse IL17A + ZJU37 group. N = 3. Experiments in Figures E and G were repeated twice. Data in Figures B, D, F, and H was calculated with ANOVA and pairwise comparison. *p < 0.05, **p < 0.01, ***p < 0.001 com pared to the vehicle group. ns indicated no significance.

Techniques: Biomarker Discovery, Inhibition, Migration, Western Blot, Comparison

C3b deposition on mouse mesangial cells. Mouse mesangial cells (MES-13) were incubated with patient serum or Factor H (FH) depleted serum with or without human FH and human Factor H-related 1*A (FHR-1*A), human Factor H-related 1*B (FHR-1*B), and/or human Factor H-related 5 (FHR-5). Cells were then stained for C3b deposition (green) and nuclei (blue). All three FHR proteins significantly increased C3b deposition as compared to FH alone, with odds increase in deposition for FHR-1*A, FHR-1*B and FHR-5 of 2.9, 11.6, and 7.3, respectively, based on 10 experimental replicates. 250μm scale bar is shown. (Logistic regression model; P < 0.001 for all).

Journal: Frontiers in Immunology

Article Title: Factor H-related 1 and heparan sulfate architecture contribute to complement dysregulation in C3 glomerulopathy

doi: 10.3389/fimmu.2025.1589674

Figure Lengend Snippet: C3b deposition on mouse mesangial cells. Mouse mesangial cells (MES-13) were incubated with patient serum or Factor H (FH) depleted serum with or without human FH and human Factor H-related 1*A (FHR-1*A), human Factor H-related 1*B (FHR-1*B), and/or human Factor H-related 5 (FHR-5). Cells were then stained for C3b deposition (green) and nuclei (blue). All three FHR proteins significantly increased C3b deposition as compared to FH alone, with odds increase in deposition for FHR-1*A, FHR-1*B and FHR-5 of 2.9, 11.6, and 7.3, respectively, based on 10 experimental replicates. 250μm scale bar is shown. (Logistic regression model; P < 0.001 for all).

Article Snippet: C3b deposition was quantified using mouse mesangial cells (MES-13)(ATCC CRL-1927; Manassas, VA) cultured on Lab-Tek II CC 2– 8 well glass chamber slides (ThermoFisher Scientific, Inc., Waltham, MA).

Techniques: Incubation, Staining

C3b deposition on mouse mesangial cells pretreated with heparinase. Mouse mesangial cells (MES-13) were treated with (A) media or (B) heparinase followed by incubation with FH-depleted serum supplemented with C3 and FH or additionally FHR-1*A, FHR-1*B or FHR-5. Cells were stained for C3b deposition (green), heparan sulfate (red), and nuclei (blue). C3b deposition was significantly increased by heparinase treatment with odds increase in deposition for FH, FHR-1*A, FHR-1*B, and FHR-5 of 2.9, 4.7, 5.5, and 1.8, respectively, based on 5 experimental replicates. 250μm scale bar is shown. (Logistic regression model; P < 0.001 for all).

Journal: Frontiers in Immunology

Article Title: Factor H-related 1 and heparan sulfate architecture contribute to complement dysregulation in C3 glomerulopathy

doi: 10.3389/fimmu.2025.1589674

Figure Lengend Snippet: C3b deposition on mouse mesangial cells pretreated with heparinase. Mouse mesangial cells (MES-13) were treated with (A) media or (B) heparinase followed by incubation with FH-depleted serum supplemented with C3 and FH or additionally FHR-1*A, FHR-1*B or FHR-5. Cells were stained for C3b deposition (green), heparan sulfate (red), and nuclei (blue). C3b deposition was significantly increased by heparinase treatment with odds increase in deposition for FH, FHR-1*A, FHR-1*B, and FHR-5 of 2.9, 4.7, 5.5, and 1.8, respectively, based on 5 experimental replicates. 250μm scale bar is shown. (Logistic regression model; P < 0.001 for all).

Techniques: Incubation, Staining

APN evaluation in vitro , with in vivo measurement of IPGTT, ITT, blood glucose and body weight in vivo. (A) C2C12 myotubes viability was assessed using the CCK8 assay . After transfection with APN-mRNA-LNP, the half maximal inhibitory concentration (IC50) was determined after 24 hours. ( B ) Relative APN Expression in differentiated Myotubes cell C2C12, (C) Relative APN Expression in Kidney mesangial cell SV40-MES13. APN gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. ( D ) APN protein expression, three separate blot analyzes of the samples were conducted. ( E ) Optical densitometry was used to semi-quantify the bands, and ImageJ (1.53t) digital imaging processing software was used for analysis. GAPDH has been used as an endogenous control for protein expression. Protein levels are represented as mean ratio values quantified from protein bands of APN versus GAPDH compared to negative control. (n = 3, Mann-Whitney U tests), Significances are shown above bars: †† P < 0.01 vs. negative control (NC), ** P < 0.01 vs. positive control (PC) “PBS” and ## P < 0.01 vs. Empty-LNP. ( F ) IPGTT with AUC of IPGTT, (G) ITT curve with AUC of ITT. The IPGTT and insulin ITT were utilized at 23 weeks old, to evaluate glucose tolerance and insulin sensitivity, respectively to depict significant changes between groups, the area under the curve (AUC) of IPGTT and ITT was calculated by the trapezoid method. Values are mean ± SD. n = 5 each group, Mann-Whitney U tests, ** P < 0.01 vs. PBS control group, NS; Non-significant. H) Blood glucose levels at different time points were evaluated by glucometer; I) Body weight at different time points. Values are mean ± SD. n = 5 each group, Mann-Whitney U tests, ** P < 0.01 vs. PBS control group and ## P < 0.01 vs. Empty-LNP group.

Journal: Aging and Disease

Article Title: Adiponectin mRNA Conjugated with Lipid Nanoparticles Specifically Targets the Pathogenesis of Type 2 Diabetes

doi: 10.14336/AD.2024.0162

Figure Lengend Snippet: APN evaluation in vitro , with in vivo measurement of IPGTT, ITT, blood glucose and body weight in vivo. (A) C2C12 myotubes viability was assessed using the CCK8 assay . After transfection with APN-mRNA-LNP, the half maximal inhibitory concentration (IC50) was determined after 24 hours. ( B ) Relative APN Expression in differentiated Myotubes cell C2C12, (C) Relative APN Expression in Kidney mesangial cell SV40-MES13. APN gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. ( D ) APN protein expression, three separate blot analyzes of the samples were conducted. ( E ) Optical densitometry was used to semi-quantify the bands, and ImageJ (1.53t) digital imaging processing software was used for analysis. GAPDH has been used as an endogenous control for protein expression. Protein levels are represented as mean ratio values quantified from protein bands of APN versus GAPDH compared to negative control. (n = 3, Mann-Whitney U tests), Significances are shown above bars: †† P < 0.01 vs. negative control (NC), ** P < 0.01 vs. positive control (PC) “PBS” and ## P < 0.01 vs. Empty-LNP. ( F ) IPGTT with AUC of IPGTT, (G) ITT curve with AUC of ITT. The IPGTT and insulin ITT were utilized at 23 weeks old, to evaluate glucose tolerance and insulin sensitivity, respectively to depict significant changes between groups, the area under the curve (AUC) of IPGTT and ITT was calculated by the trapezoid method. Values are mean ± SD. n = 5 each group, Mann-Whitney U tests, ** P < 0.01 vs. PBS control group, NS; Non-significant. H) Blood glucose levels at different time points were evaluated by glucometer; I) Body weight at different time points. Values are mean ± SD. n = 5 each group, Mann-Whitney U tests, ** P < 0.01 vs. PBS control group and ## P < 0.01 vs. Empty-LNP group.

Article Snippet: SV40 MES13: Mouse mesangial glomerulus SV40-MES13 kidney cells were obtained from the American Type Culture Collection (ATCC CRL-1927) (Manassas, VA).

Techniques: In Vitro, In Vivo, CCK-8 Assay, Transfection, Concentration Assay, Expressing, Gene Expression, Imaging, Software, Control, Negative Control, MANN-WHITNEY, Positive Control

Relative gene expression in different tissues . ( A ) Glut-4 gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. ( B ) DGKd gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, and Kidney. ( C ) PKCe gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. Gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro. Values are mean ± SD. n = 9, student t-test. In vivo, a total of five biological samples (n = 5) were collected. Of each sample, three replicates were performed technically. ( D ) Glut-4, DGKd, and PKCe protein expression, three separate blot analysis of the samples were conducted. ( E ) Optical densitometry was used to semi-quantify the bands, and ImageJ (1.53t) digital imaging processing software was used for analysis. GAPDH has been used as an endogenous control for protein expression. Protein levels are represented as mean ratio values quantified from protein bands of the targeted protein versus GAPDH compared to negative control. (n = 3, Mann-Whitney U tests), Significances are shown above bars: †† P < 0.01 vs. negative control (NC), ** P < 0.01 vs. positive control (PC) “PBS” and ## P < 0.01 vs. Empty-LNP.

Journal: Aging and Disease

Article Title: Adiponectin mRNA Conjugated with Lipid Nanoparticles Specifically Targets the Pathogenesis of Type 2 Diabetes

doi: 10.14336/AD.2024.0162

Figure Lengend Snippet: Relative gene expression in different tissues . ( A ) Glut-4 gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. ( B ) DGKd gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, and Kidney. ( C ) PKCe gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. Gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro. Values are mean ± SD. n = 9, student t-test. In vivo, a total of five biological samples (n = 5) were collected. Of each sample, three replicates were performed technically. ( D ) Glut-4, DGKd, and PKCe protein expression, three separate blot analysis of the samples were conducted. ( E ) Optical densitometry was used to semi-quantify the bands, and ImageJ (1.53t) digital imaging processing software was used for analysis. GAPDH has been used as an endogenous control for protein expression. Protein levels are represented as mean ratio values quantified from protein bands of the targeted protein versus GAPDH compared to negative control. (n = 3, Mann-Whitney U tests), Significances are shown above bars: †† P < 0.01 vs. negative control (NC), ** P < 0.01 vs. positive control (PC) “PBS” and ## P < 0.01 vs. Empty-LNP.

Article Snippet: SV40 MES13: Mouse mesangial glomerulus SV40-MES13 kidney cells were obtained from the American Type Culture Collection (ATCC CRL-1927) (Manassas, VA).

Techniques: Gene Expression, Muscles, In Vitro, In Vivo, Expressing, Imaging, Software, Control, Negative Control, MANN-WHITNEY, Positive Control

Relative IR expression in different studied tissues . ( A ) Differentiated Myotubes cell C2C12; (B) Kidney mesangial cell SV40-MES13; (C) Skeletal muscles; (D) Liver; (E) Kidney. IR gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. In vivo , a total of five biological samples (n = 5, student t-test) were collected. Of each sample, three replicates were performed technically. Significances are shown above bars.

Journal: Aging and Disease

Article Title: Adiponectin mRNA Conjugated with Lipid Nanoparticles Specifically Targets the Pathogenesis of Type 2 Diabetes

doi: 10.14336/AD.2024.0162

Figure Lengend Snippet: Relative IR expression in different studied tissues . ( A ) Differentiated Myotubes cell C2C12; (B) Kidney mesangial cell SV40-MES13; (C) Skeletal muscles; (D) Liver; (E) Kidney. IR gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. In vivo , a total of five biological samples (n = 5, student t-test) were collected. Of each sample, three replicates were performed technically. Significances are shown above bars.

Article Snippet: SV40 MES13: Mouse mesangial glomerulus SV40-MES13 kidney cells were obtained from the American Type Culture Collection (ATCC CRL-1927) (Manassas, VA).

Techniques: Expressing, Muscles, Gene Expression, In Vitro, In Vivo

EGFR expression in vitro and in vivo . (A) Relative EGFR Expression in different Differentiated Myotubes cell C2C12; (B) Kidney mesangial cell SV40-MES13; (C) kidney. EGFR gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. In vivo , a total of five biological samples (n = 5, student t-test) were collected. Of each sample, three replicates were performed technically. Significances are shown above bars. ( D ) Overall correlation between APN and EGFR gene expression in kidney. ( E ) Light microscopic appearance in kidney tissue section of the studied groups of DIO mice, (yellow arrow) indicates Hyperplasia of partial cells in the Bowmans capsule of the glomeruli and (Black arrow) indicates mild diffuse mesangial matrix expansion (PAS stain, 1000X). A total of five biological samples (n = 5, student t-test) were collected; three sections were taken from each sample. Scale bar= 50 μm.

Journal: Aging and Disease

Article Title: Adiponectin mRNA Conjugated with Lipid Nanoparticles Specifically Targets the Pathogenesis of Type 2 Diabetes

doi: 10.14336/AD.2024.0162

Figure Lengend Snippet: EGFR expression in vitro and in vivo . (A) Relative EGFR Expression in different Differentiated Myotubes cell C2C12; (B) Kidney mesangial cell SV40-MES13; (C) kidney. EGFR gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Fold-change values less than one indicate downregulation, whereas values greater than one indicate upregulation. Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. In vivo , a total of five biological samples (n = 5, student t-test) were collected. Of each sample, three replicates were performed technically. Significances are shown above bars. ( D ) Overall correlation between APN and EGFR gene expression in kidney. ( E ) Light microscopic appearance in kidney tissue section of the studied groups of DIO mice, (yellow arrow) indicates Hyperplasia of partial cells in the Bowmans capsule of the glomeruli and (Black arrow) indicates mild diffuse mesangial matrix expansion (PAS stain, 1000X). A total of five biological samples (n = 5, student t-test) were collected; three sections were taken from each sample. Scale bar= 50 μm.

Article Snippet: SV40 MES13: Mouse mesangial glomerulus SV40-MES13 kidney cells were obtained from the American Type Culture Collection (ATCC CRL-1927) (Manassas, VA).

Techniques: Expressing, In Vitro, In Vivo, Gene Expression, Staining

Relative gene expression of the pro-inflammatory cytokines . ( A ) TNFa gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. ( B ) IL-6 gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. ( C ) IL-1b gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. Gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. In vivo , a total of five biological samples (n = 5) were collected. Of each sample, three replicates were performed technically. ( D ) Glut-4 TNFa and IL-6 protein expression, three separate blot analysis of the samples were conducted. ( E ) Optical densitometry was used to semi-quantify the bands, and ImageJ (1.53t) digital imaging processing software was used for analysis. GAPDH has been used as an endogenous control for protein expression. Protein levels are represented as mean ratio values quantified from protein bands of the targeted protein versus GAPDH compared to negative control. (n = 3, Mann-Whitney U tests), Significances are shown above bars: †† P < 0.01 vs. negative control (NC), ** P < 0.01 vs. positive control (PC) “PBS” and ## P < 0.01 vs. Empty-LNP.

Journal: Aging and Disease

Article Title: Adiponectin mRNA Conjugated with Lipid Nanoparticles Specifically Targets the Pathogenesis of Type 2 Diabetes

doi: 10.14336/AD.2024.0162

Figure Lengend Snippet: Relative gene expression of the pro-inflammatory cytokines . ( A ) TNFa gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. ( B ) IL-6 gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. ( C ) IL-1b gene in differentiated myotubes cell C2C12, Kidney mesangial cell SV40-MES13, Skeletal muscles, Liver, Kidney, White fats, and Brown fats. Gene expression was calculated based on the Law of Fold-change, which is 2 -ΔΔCT . Both biological and technical triplicates were performed in vitro . Values are mean ± SD. n = 9, student t-test. In vivo , a total of five biological samples (n = 5) were collected. Of each sample, three replicates were performed technically. ( D ) Glut-4 TNFa and IL-6 protein expression, three separate blot analysis of the samples were conducted. ( E ) Optical densitometry was used to semi-quantify the bands, and ImageJ (1.53t) digital imaging processing software was used for analysis. GAPDH has been used as an endogenous control for protein expression. Protein levels are represented as mean ratio values quantified from protein bands of the targeted protein versus GAPDH compared to negative control. (n = 3, Mann-Whitney U tests), Significances are shown above bars: †† P < 0.01 vs. negative control (NC), ** P < 0.01 vs. positive control (PC) “PBS” and ## P < 0.01 vs. Empty-LNP.

Article Snippet: SV40 MES13: Mouse mesangial glomerulus SV40-MES13 kidney cells were obtained from the American Type Culture Collection (ATCC CRL-1927) (Manassas, VA).

Techniques: Gene Expression, Muscles, In Vitro, In Vivo, Expressing, Imaging, Software, Control, Negative Control, MANN-WHITNEY, Positive Control

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