Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation exacerbated mortality in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, 50, or 100 µM AG490, followed by co-treatment with 0 or 10 µM MeHg. Cytotoxicity (LDH release) was measured using LDH at 3 h ( A ), 6 h ( B ), and 24 h ( C ) of MeHg addition. Cell viability was assessed using MTT at 3 h ( D ), 6 h ( E ), and 24 h ( F ) of MeHg addition. Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation increases oxidative stress in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, 50, or 100 µM AG490 ( A - E ) or with 0, 3, or 30 µM C188-9 ( F - J ), followed by co-treatment with 0 or 10 µM MeHg. ROS production was measured using CM-H2DCFDA probe after MeHg addition at 30 min ( A , F ), 1 h ( B , G ), 3 h ( C , H ), 6 h ( D , I ), and 24 h ( E , J ). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation increases mitochondrial oxidative stress in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, 50, or 100 µM AG490 ( A - D ) or with 0, 3, or 30 µM C188-9 ( F - H ), followed by co-treatment with 0 or 10 µM MeHg. Mitochondrial superoxide production was assessed using mitoSOX fluorescent staining, after MeHg addition at 30 min ( A , E ), 1 h ( B , F ), 3 h ( C , G ), and 6 h ( D , H ). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Staining, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation exacerbates GSH depletion in C8-D1A astrocytic cells. Cells were pretreated with 0 or 100 µM AG490 ( A , B , C ) or with 0 or 30 µM C188-9 ( D , E , F ), followed by co-treatment with 0 or 10 µM MeHg. Total GSH ( A , D ) reduced GSH ( B , C ), and GSH/GSSG ratio were measured at 24 h after adding MeHg. Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: STAT3 inhibition induces aberrant antioxidant enzyme expression in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, or 100 µM AG490 or with 0 or 30 µM C188-9, followed by 24 h of co-treatment with 0 or 10 µM MeHg. Protein levels of phosphorylated STAT3 to total STAT3 ratio (pSTAT3/STAT3) ( A , E ), HO-1 ( B , F ), and Nrf2 ( C , G ) were measured by western blot. Representative densitometry images are shown in ( D , H ). Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. P < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Expressing, Western Blot, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: STAT3 inhibition induces aberrant antioxidant gene expression in C8-D1A astrocytic cells. Cells were pretreated with 0, 10, or 100 µM AG490 or with 0 or 30 µM C188-9, followed by 24 h of co-treatment with 0 or 10 µM MeHg. Gene expression of Stat3 ( A , C ) and Socs3 ( B , D ) at 3 h, and Stat3 ( E , I ), Socs3 ( F , J ), Hmox1 ( G , K ), and Slc7a11 ( H , L ) at 24 h was measured by qPCR. Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Gene Expression, Transformation Assay

Journal: Neurochemical Research

Article Title: JAK2/STAT3 Signaling Pathway Modulates Acute Methylmercury Toxicity in the Mouse Astrocyte C8-D1A Cell Line

doi: 10.1007/s11064-025-04507-7

Figure Lengend Snippet: STAT3 inhibition did not prevent MeHg IL-6 release in C8-D1A astrocytic cells. Cells were pretreated with 0, 10 or 100 µM AG490 ( A , B ) or with 0, 3, or 30 µM C188-9 ( C , D ), followed by co-treatment with 0 or 10 µM MeHg. Il- 6 mRNA expression ( A , C ) was measured by qPCR at 3 h after adding MeHg. IL-6 release into the medium ( B , D ) was measured at 16 h after adding MeHg Data are presented as mean ± SD. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s post-hoc analysis. When data did not meet the assumptions of normality, a logarithmic or square root transformation was employed before conducting the two-way ANOVA. p < 0.05 was considered statistically significant. * denotes a significant difference

Article Snippet: Cells were pre-treated for 1 h with JAK2 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10 μM MeHg at specified times.

Techniques: Inhibition, Expressing, Transformation Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Evidence for a JAK2/STAT3 proinflammatory and vasculogenic mimicry interrelated molecular signature in adipocyte-derived mesenchymal stromal/stem cells

doi: 10.1186/s12964-025-02298-6

Figure Lengend Snippet: Effect of JAK2/STAT3 signaling pathway inhibition on the formation of 3D capillary-like structures in ADMSCs. ADMSCs were trypsinized and seeded on top of a Cultrex layer as described in the Methods section. ( A ) Representative phase contrast images were taken to monitor the formation of capillary-like structures (upper panels) and were analyzed via Wimasis (lower panels). ( B ) Quantification from Wimasis analysis of the total loops and branching points over time. ( C ) Representative phase contrast images were taken to monitor the inhibition of capillary-like structure formation by increasing concentrations of AG490 after 5 h (upper panels). The structures were analyzed via Wimasis (lower panels). ( D ) Representative phase contrast pictures were taken to monitor the inhibition of capillary-like structure formation by RNA-mediated interference via siSTAT3 in transiently transfected ADMSCs after 5 h (upper panels) compared with that in control cells transiently transfected with a scrambled sequence (siScr); the structures were analyzed via Wimasis (lower panels). ( E ) Quantification from Wimasis analysis of the total loops and of branching points at 10 µM AG490 (black bars) and upon siSTAT3-transfection (gray bars) are shown as percentages of their respective controls (0.1% DMSO and siScr). ( F ) Relative gene expression of STAT3 after pharmacological inhibition with 10 µM AG490 (black bars) and upon transient interference with siSTAT3 (gray bars). The VM data are representative of three independent experiments, and the transfection data are representative of two independent experiments

Article Snippet: Tyrphostin AG490 (133550-30-8) was obtained from Calbiochem (La Jolla, CA, USA).

Techniques: Inhibition, Transfection, Control, Sequencing, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Evidence for a JAK2/STAT3 proinflammatory and vasculogenic mimicry interrelated molecular signature in adipocyte-derived mesenchymal stromal/stem cells

doi: 10.1186/s12964-025-02298-6

Figure Lengend Snippet: Effect of pharmacological inhibition of the JAK2/STAT3 signaling pathway on the chemotactic migration capacity of ADMSCs. Chemotactic real-time cell migration was performed as described in the Methods section. ( A ) Relative cell migration in serum-supplemented or serum-free media was monitored over a 3-hour period. ( B ) A representative profile of the effects of the indicated increasing concentrations of AG490 on relative cell migration was compared to that of the vehicle (0.1% DMSO) over 3 h. ( C ) Quantification of the inhibitory effect of AG490 concentration on the chemotactic response. All the data are representative of two independent experiments, each performed in triplicate

Article Snippet: Tyrphostin AG490 (133550-30-8) was obtained from Calbiochem (La Jolla, CA, USA).

Techniques: Inhibition, Migration, Concentration Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Evidence for a JAK2/STAT3 proinflammatory and vasculogenic mimicry interrelated molecular signature in adipocyte-derived mesenchymal stromal/stem cells

doi: 10.1186/s12964-025-02298-6

Figure Lengend Snippet: Transcriptomic analysis of the inflammatory molecular signature induced upon capillary-like structure formation. ADMSCs were seeded on top of Cultrex dishes and left to form 3D capillary-like structures for 5 h in the presence or absence of 10 µM AG490 (the control consisted of vehicle-treated 2D monolayers). Total RNA was extracted, and gene expression was assessed via an RT 2 Profiler PCR array for human inflammatory cytokines and receptors as described in the Methods section. ( A ) Gene expression levels of upregulated genes (green arrow) and downregulated genes (red arrow) are represented as the fold regulation between the control 2D monolayers and VM-forming ADMSCs, with a cutoff value greater than or equal to two. ( B ) Number of VM-induced genes and extent of their AG490-mediated inhibition of expression. ( C ) Protein‒protein interaction network of the 10 most VM-induced genes whose expression was inhibited by AG490, as retrieved with STRING. ( D ) Number of VM-downregulated genes and extent of their AG490-mediated inhibition of expression. ( E ) Protein‒protein interaction network of the 10 most VM-downregulated genes whose expression was also inhibited by AG490, as determined with STRING

Article Snippet: Tyrphostin AG490 (133550-30-8) was obtained from Calbiochem (La Jolla, CA, USA).

Techniques: Control, Gene Expression, Inhibition, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Evidence for a JAK2/STAT3 proinflammatory and vasculogenic mimicry interrelated molecular signature in adipocyte-derived mesenchymal stromal/stem cells

doi: 10.1186/s12964-025-02298-6

Figure Lengend Snippet: Effects of AG490 on inflammation and invasion markers related to capillary-like structure formation. ADMSC response to priming on Cultrex for 5 h for capillary-like structure formation in the presence or absence of 10 µM AG490 (control, 2D monolayers). Total RNA was extracted, and gene expression was assessed via RT‒qPCR, as described in the Methods section. Relative gene expression of inflammation and invasion markers was compared between the control (vehicle-treated 2D monolayers, white bars) and VM-forming ADMSCs in the presence (3D + AG490; gray bars) or absence (vehicle-treated 3D; black bars) of 10 µM AG490. The results are presented as percentages of VM-related gene expression levels. The data are representative of two to three independent experiments performed in triplicate, except for the AG490 data, which are representative of one to two independent experiments performed in triplicate

Article Snippet: Tyrphostin AG490 (133550-30-8) was obtained from Calbiochem (La Jolla, CA, USA).

Techniques: Control, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Evidence for a JAK2/STAT3 proinflammatory and vasculogenic mimicry interrelated molecular signature in adipocyte-derived mesenchymal stromal/stem cells

doi: 10.1186/s12964-025-02298-6

Figure Lengend Snippet: Involvement of the JAK2/STAT3 signaling pathway in the TNFα-induced inflammatory signature in ADMSCs. ( A ) Monolayer ADMSC culture response to increasing TNFα concentrations for 24 h. COX-2, phospho-STAT3 (pSTAT3) and total STAT3 protein lysates were immunoblotted, with GAPDH serving as a loading control. ( B ) Densitometry analysis of COX-2 and pSTAT3 expression, represented as the percentage of the respective maximal effect of TNFα. ( C ) ADMSC 2D monolayer cultures treated with 30 ng/mL TNFα in the presence of the indicated increasing concentrations of AG490 for 24 h (vehicle, 0.1% DMSO). COX-2, pSTAT3 and STAT3 protein lysates were immunoblotted, with GAPDH serving as a loading control. ( D ) Densitometry analysis of TNFα-induced COX-2/GAPDH inhibition, represented as the percentage of the maximal effect of TNFα (labeled COX-2; left panel), and of TNFα-induced pSTAT3/STAT3 (labeled pSTAT3; right panel). ( E ) Monolayer-ADMSC cultures subjected to 30 ng/mL TNFα in the presence or absence of 10 µM AG490 for 24 h (the control was 0.1% DMSO). Modulation profiles of inflammation and invasion gene markers by TNFα (black bars) and by AG490 (gray bars) were measured via RT‒qPCR. All the data are representative of three independent experiments

Article Snippet: Tyrphostin AG490 (133550-30-8) was obtained from Calbiochem (La Jolla, CA, USA).

Techniques: Control, Expressing, Inhibition, Labeling

Journal: Cell Communication and Signaling : CCS

Article Title: Evidence for a JAK2/STAT3 proinflammatory and vasculogenic mimicry interrelated molecular signature in adipocyte-derived mesenchymal stromal/stem cells

doi: 10.1186/s12964-025-02298-6

Figure Lengend Snippet: Implication of JAK2/STAT3 in the inflammatory molecular signature of ADMSCs stimulated with TNFα. ADMSCs were treated with 30 ng/mL TNFα in the presence or absence of 10 µM AG490 for 24 h (the control was treated with 0.1% DMSO) to perform a RT 2 PCR array for human inflammatory cytokines and receptors, as described in the Methods section. ( A ) Gene expression of upregulated genes (green arrow) and downregulated genes (red arrow) is represented as the fold regulation between the control and TNFα-treated cells, with a cutoff value greater than or equal to two. ( B ) Number of TNFα-induced genes and extent to which AG490 inhibited their expression. ( C ) Protein‒protein interaction network of the 10 TNFα-induced genes whose expression was inhibited the most by AG490, as determined via the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING). ( D ) Number of TNFα-downregulated genes and extent to which AG490 inhibited their expression. ( E ) Protein‒protein interaction network of the 10 genes most downregulated by TNFα that were inhibited by AG490, as determined with STRING. The data are representative of one of two independent experiments

Article Snippet: Tyrphostin AG490 (133550-30-8) was obtained from Calbiochem (La Jolla, CA, USA).

Techniques: Control, Gene Expression, Expressing

Journal: International Journal of Molecular Sciences

Article Title: SOCS1 Inhibits IL-6-Induced CD155 Overexpression in Lung Adenocarcinoma

doi: 10.3390/ijms252212141

Figure Lengend Snippet: Effect of IL-6 stimulation on CD155 expression on lung adenocarcinoma cell lines. ( A ) IL-6 stimulation induced CD155 mRNA expression in the tested cell lines. ( B ) Upon IL-6 stimulation, CD155 protein expression increased in A-549 and NCI-H1437 cells, but no change was detected in the advanced stages 3B1A and NCI-H1573 cell lines. ( C ) Results show that CD155 concentration was augmented only in the supernatant of the IL-6-stimulated A-549 cell line. ( D ) Addition of the JAK2 inhibitor AG490 prevents IL-6-induced CD155 overexpression in the A-549 and NCI-H1437 cell lines. ( A , C ) are shown as mean + SD. * p < 0.05, ** p < 0.01.

Article Snippet: In addition, to confirm that the observed effect was IL-6 dependent, the JAK2 inhibitor, Tyrphostin AG490 (Cat. No. S1143; Selleckchem, Houston, TX, USA), was added at a concentration of 15 μM prior to IL-6 addition.

Techniques: Expressing, Concentration Assay, Over Expression

Journal: bioRxiv

Article Title: JAK2/STAT3 signaling pathway mediates methylmercury toxicity in mouse astrocyte neuronal C8-D1A cell line

doi: 10.1101/2024.07.13.603400

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation enhanced MeHg toxicity in C8-D1A cells. Astrocytic C8-D1A cells were treated with AG490 (0, 10, 50, or 100µM) for one hour followed by co-exposure to MeHg (0 or 10µM) for 3 hours (A and D), 6 hours (B and E), and 24 hours (C and F). Cytotoxicity was measured using the LDH assay (A, B, and C), and viability was evaluated using the MTT assay (D, E, and F). Data are represented as mean ± SD. Data were analyzed by two-way ANOVA followed by Tukey’s post hoc test. * denotes statistically significant compared to the respective control (p < 0.05).

Article Snippet: Cells were pre-treated for 1h with STAT3 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Lactate Dehydrogenase Assay, MTT Assay, Control

Journal: bioRxiv

Article Title: JAK2/STAT3 signaling pathway mediates methylmercury toxicity in mouse astrocyte neuronal C8-D1A cell line

doi: 10.1101/2024.07.13.603400

Figure Lengend Snippet: Blocking STAT3 phosphorylation enhances MeHg-induced intracellular ROS production in C8-D1A cells. Astrocytic C8-D1A cells were treated with AG490 (0, 10, 50, or 100µM) for one hour followed by co-exposure to MeHg (0 or 10µM) for 0 hours (A), 30 min (B), 1 hours (C), 3 hours (D), 6 hours (E), and 24 hours (F). Intracellular ROS was measured using the CM-H 2 DCFDA fluorescent probe. Data are represented as percentage of control as mean ± SD. Data were analyzed by two-way ANOVA followed by Tukey’s post hoc test. * denotes statistically significant compared to the respective control (p < 0.05).

Article Snippet: Cells were pre-treated for 1h with STAT3 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10μM MeHg at specified times.

Techniques: Blocking Assay, Phospho-proteomics, Control

Journal: bioRxiv

Article Title: JAK2/STAT3 signaling pathway mediates methylmercury toxicity in mouse astrocyte neuronal C8-D1A cell line

doi: 10.1101/2024.07.13.603400

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation aggravates MeHg-induced mitochondrial ROS production in C8-D1A cells. Astrocytic C8-D1A cells were treated with AG490 (0, 10, 50, or 100µM) for one hour followed by co-exposure to MeHg (0 or 10µM) for 0 hours (A), 30 min (B), 1 hours (C), 3 hours (D), and 6 hours (E). Mitochondrial ROS was measured using the mitoSOX fluorescent probe. Data are represented as percentage of control as mean ± SD. Data were analyzed by two-way ANOVA followed by Tukey’s post hoc test. * denotes statistically significant compared to the respective control (p < 0.05).

Article Snippet: Cells were pre-treated for 1h with STAT3 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Control

Journal: bioRxiv

Article Title: JAK2/STAT3 signaling pathway mediates methylmercury toxicity in mouse astrocyte neuronal C8-D1A cell line

doi: 10.1101/2024.07.13.603400

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation exacerbates MeHg-induced HO1 protein expression in C8-D1A cells. Astrocytic C8-D1A cells were treated with AG490 (0, 10, 50, or 100µM) for one hour followed by co-exposure to MeHg (0 or 10µM) for 1 hour. Western blot analysis was performed to measure (A), pSTAT3, (B), STAT3, (C) pSTAT3/STAT3 ratio, and (D) HO1 protein expression levels, quantified by densitometry. (E), Representative western blot of the total and phosphorylated proteins are shown. Data are presented as percentage of control as mean ± SD. Data were analyzed by two-way ANOVA followed by Tukey’s post hoc test. * denotes statistically significant compared to the respective control (p < 0.05).

Article Snippet: Cells were pre-treated for 1h with STAT3 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: JAK2/STAT3 signaling pathway mediates methylmercury toxicity in mouse astrocyte neuronal C8-D1A cell line

doi: 10.1101/2024.07.13.603400

Figure Lengend Snippet: Inhibition of STAT3 phosphorylation exacerbates MeHg-induced HO1 protein expression in C8-D1A cells. Astrocytic C8-D1A cells were treated with AG490 (0, 10, 50, or 100µM) for one hour followed by co-exposure to MeHg (0 or 10µM) for 3 hours. Western blot analysis was performed to measure (A), pSTAT3, (B), STAT3, (C) pSTAT3/STAT3 ratio, (D) HO1, (F) Nrf2 protein expression levels, quantified by densitometry. (F), Representative western blot of the total and phosphorylated proteins are shown. Data are presented as percentage of control as mean ± SD. Data were analyzed by two-way ANOVA followed by Tukey’s post hoc test. * denotes statistically significant compared to the respective control (p < 0.05).

Article Snippet: Cells were pre-treated for 1h with STAT3 inhibitor AG490 (Tyrphostin B42 or 2-cyano-3-(3,4-dihydroxyphenyl)-N-(phenylmethyl)-2-propenamideor), Santa Cruz Biotechnology, sc-202046) at 0, 10, 50, or 100 μM, followed by co-treatment with 0 or 10μM MeHg at specified times.

Techniques: Inhibition, Phospho-proteomics, Expressing, Western Blot, Control