sb202190 fhpi  (Selleck Chemicals)


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    Name:
    SB202190
    Description:
    SB202190 FHPI is a potent p38 MAPK inhibitor targeting p38α β with IC50 of 50 nM 100 nM in cell free assays sometimes used instead of SB 203580 to investigate potential roles for SAPK2a p38 in vivo SB202190 inhibits endothelial cell apoptosis via induction of autophagy and heme oxygenase 1 SB202190 significantly suppresses Erastin dependent ferroptosis
    Catalog Number:
    s1077-25mg
    Price:
    147.0
    Size:
    25mg
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    Structured Review

    Selleck Chemicals sb202190 fhpi
    SB202190
    SB202190 FHPI is a potent p38 MAPK inhibitor targeting p38α β with IC50 of 50 nM 100 nM in cell free assays sometimes used instead of SB 203580 to investigate potential roles for SAPK2a p38 in vivo SB202190 inhibits endothelial cell apoptosis via induction of autophagy and heme oxygenase 1 SB202190 significantly suppresses Erastin dependent ferroptosis
    https://www.bioz.com/result/sb202190 fhpi/product/Selleck Chemicals
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sb202190 fhpi - by Bioz Stars, 2021-03
    95/100 stars

    Images

    1) Product Images from "A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway"

    Article Title: A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway

    Journal: Journal of Developmental Biology

    doi: 10.3390/jdb8030012

    p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.
    Figure Legend Snippet: p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.

    Techniques Used: Concentration Assay

    2) Product Images from "A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway"

    Article Title: A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway

    Journal: Journal of Developmental Biology

    doi: 10.3390/jdb8030012

    p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.
    Figure Legend Snippet: p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.

    Techniques Used: Concentration Assay

    3) Product Images from "Identification of SYK inhibitor, R406 as a novel senolytic agent"

    Article Title: Identification of SYK inhibitor, R406 as a novel senolytic agent

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.103135

    R406 inhibits phosphorylation of p38 MAPK in senescent HDFs more than non-senescent HDFs. ( A , B ) Senescent (S) and non-senescent (NS) HDFs were treated with DMSO, R406 (10 μM), and ABT263 (1 μM) for one day and western blot assays using anti-MAPKs antibodies were conducted. ( C , D ) Senescent HDFs were respectively treated with DMSO, R406 (10 μM), and SB202190 (p38 inhibitor, 50 μM) for one day. Then, western blot assays were conducted to determine triggering apoptosis by caspase cleavage. *p
    Figure Legend Snippet: R406 inhibits phosphorylation of p38 MAPK in senescent HDFs more than non-senescent HDFs. ( A , B ) Senescent (S) and non-senescent (NS) HDFs were treated with DMSO, R406 (10 μM), and ABT263 (1 μM) for one day and western blot assays using anti-MAPKs antibodies were conducted. ( C , D ) Senescent HDFs were respectively treated with DMSO, R406 (10 μM), and SB202190 (p38 inhibitor, 50 μM) for one day. Then, western blot assays were conducted to determine triggering apoptosis by caspase cleavage. *p

    Techniques Used: Western Blot

    R406 induces senolytic effects via blocking the phosphorylation of p38 MAPK and FAK simultaneously. ( A ) Senescent (S) and non-senescent (NS) HDFs were respectively treated with DMSO, R406 (10 μM), PF562271 (5 μM), and SB202190 (50 μM) for one day and then western blot assay with anti-caspase-9 antibody was conducted to determine apoptosis. ( B ) Proposed mechanism of senolytic effect by R406. R406-induced cell death is mediated by inhibiting FAK and p38 activity as well as increasing ROS. *p
    Figure Legend Snippet: R406 induces senolytic effects via blocking the phosphorylation of p38 MAPK and FAK simultaneously. ( A ) Senescent (S) and non-senescent (NS) HDFs were respectively treated with DMSO, R406 (10 μM), PF562271 (5 μM), and SB202190 (50 μM) for one day and then western blot assay with anti-caspase-9 antibody was conducted to determine apoptosis. ( B ) Proposed mechanism of senolytic effect by R406. R406-induced cell death is mediated by inhibiting FAK and p38 activity as well as increasing ROS. *p

    Techniques Used: Blocking Assay, Western Blot, Activity Assay

    4) Product Images from "Pyrimethamine Elicits Antitumor Effects on Prostate Cancer by Inhibiting the p38-NF-κB Pathway"

    Article Title: Pyrimethamine Elicits Antitumor Effects on Prostate Cancer by Inhibiting the p38-NF-κB Pathway

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2020.00758

    Pyrimethamine inhibits proliferation and promotes apoptosis in CRPC cells through the inhibition of p38 MAPK. (A, B) DU145 and PC3 cells were treated with pyrimethamine or SB202190 (10 μM) for 24 h and then incubated with TNF-a (10 ng/ml) for 24 h, and quantification of percentages in cell cycle phases was analyzed by flow cytometry. (C, D) Mean percentage of cells ± SD was shown (DU145 and PC3). **:P
    Figure Legend Snippet: Pyrimethamine inhibits proliferation and promotes apoptosis in CRPC cells through the inhibition of p38 MAPK. (A, B) DU145 and PC3 cells were treated with pyrimethamine or SB202190 (10 μM) for 24 h and then incubated with TNF-a (10 ng/ml) for 24 h, and quantification of percentages in cell cycle phases was analyzed by flow cytometry. (C, D) Mean percentage of cells ± SD was shown (DU145 and PC3). **:P

    Techniques Used: Inhibition, Incubation, Flow Cytometry

    Pyrimethamine regulates the p65/NF- k B pathway in CRPC cells through p38 MAPK inhibition. (A, B) The western blot analysis of the expression of p38, pp38, and pp65 after treatment of 0, 32, and 100 μM pyrimethamine for 24 h in DU145 and PC3 cell lines. The ratio of pp38/p38 was normalized to GAPDH. (C, D) DU145 and PC3 cells were treated for 24 h with pyrimethamine or SB202190 (10 μM) and then treated for 6 h in the absence or presence of TNF-a (10 ng/ml). Total cell lysates were analyzed by western blot with antibodies against p38, pp38, and pP65. (E) A consolidated model that illustrates a plausible sequence for the mechanism by which pyrimethamine elicits antitumor effects in prostate cancer.
    Figure Legend Snippet: Pyrimethamine regulates the p65/NF- k B pathway in CRPC cells through p38 MAPK inhibition. (A, B) The western blot analysis of the expression of p38, pp38, and pp65 after treatment of 0, 32, and 100 μM pyrimethamine for 24 h in DU145 and PC3 cell lines. The ratio of pp38/p38 was normalized to GAPDH. (C, D) DU145 and PC3 cells were treated for 24 h with pyrimethamine or SB202190 (10 μM) and then treated for 6 h in the absence or presence of TNF-a (10 ng/ml). Total cell lysates were analyzed by western blot with antibodies against p38, pp38, and pP65. (E) A consolidated model that illustrates a plausible sequence for the mechanism by which pyrimethamine elicits antitumor effects in prostate cancer.

    Techniques Used: Inhibition, Western Blot, Expressing, Sequencing

    5) Product Images from "A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway"

    Article Title: A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway

    Journal: Journal of Developmental Biology

    doi: 10.3390/jdb8030012

    p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.
    Figure Legend Snippet: p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.

    Techniques Used: Concentration Assay

    6) Product Images from "SUMOylation is essential for Sirt2 tumor-suppressor function in neuroblastoma"

    Article Title: SUMOylation is essential for Sirt2 tumor-suppressor function in neuroblastoma

    Journal: Neoplasia (New York, N.Y.)

    doi: 10.1016/j.neo.2020.11.013

    Sirt2-SUMOylation mainly suppresses P38-mTORC2-AKT signaling in neuroblastoma cells. (A) SH-SY5Y cells infected with lentiviral expressing shRNA targeting to Sirt2 3’-UTR or control were starved for 2 h and then stimulated by complete culture medium for 5 min. Cell lysis was collected and resolved by SDS-PAGE for western-blot analysis. ( B–E) SH-SY5Y cells infected with lentiviral expressing Sirt2 shRNA or control were treated with mTORC1 inhibitor Rapamycin (20 ug/mL), AKT inhibitor LY294002 (50 μM), P38 inhibitor SB202190 (20 μM), pp242 (0.5 or 1 μM) or dimethylsulfoxide (DMSO) as a control for 60 min. Cell lysis was performed and visualized by SDS-PAGE for western-blot analysis (B and D) . Protein levels were quantified by Image J software and showed as a graph (C and E) . ( F) A snapshot of the relationship of P38-mTOR-AKT signal mediated by Sirt2. ( G) The homemade lentivector contained CMV promoter for overexpressing foreign gene, H1 promoter for expressing shRNA and EF1a promoter for Puromycin and EGFP expression. The vector map was drawn by using DNAMAN software. ( H and I) Cell lines of SH-SY5Y harboring silenced endogenous Sirt2 with or without expressing exogenous wild-type Sirt2 or its mutants were harvested for western blot analysis (H) . Protein levels were quantified by Image J software and are shown as a graph (I) . All the western-blot analysis was repeated at least 3 times. * P
    Figure Legend Snippet: Sirt2-SUMOylation mainly suppresses P38-mTORC2-AKT signaling in neuroblastoma cells. (A) SH-SY5Y cells infected with lentiviral expressing shRNA targeting to Sirt2 3’-UTR or control were starved for 2 h and then stimulated by complete culture medium for 5 min. Cell lysis was collected and resolved by SDS-PAGE for western-blot analysis. ( B–E) SH-SY5Y cells infected with lentiviral expressing Sirt2 shRNA or control were treated with mTORC1 inhibitor Rapamycin (20 ug/mL), AKT inhibitor LY294002 (50 μM), P38 inhibitor SB202190 (20 μM), pp242 (0.5 or 1 μM) or dimethylsulfoxide (DMSO) as a control for 60 min. Cell lysis was performed and visualized by SDS-PAGE for western-blot analysis (B and D) . Protein levels were quantified by Image J software and showed as a graph (C and E) . ( F) A snapshot of the relationship of P38-mTOR-AKT signal mediated by Sirt2. ( G) The homemade lentivector contained CMV promoter for overexpressing foreign gene, H1 promoter for expressing shRNA and EF1a promoter for Puromycin and EGFP expression. The vector map was drawn by using DNAMAN software. ( H and I) Cell lines of SH-SY5Y harboring silenced endogenous Sirt2 with or without expressing exogenous wild-type Sirt2 or its mutants were harvested for western blot analysis (H) . Protein levels were quantified by Image J software and are shown as a graph (I) . All the western-blot analysis was repeated at least 3 times. * P

    Techniques Used: Infection, Expressing, shRNA, Lysis, SDS Page, Western Blot, Software, Plasmid Preparation

    7) Product Images from "A stress response p38 MAP kinase inhibitor SB202190 promoted TFEB/TFE3-dependent autophagy and lysosomal biogenesis independent of p38"

    Article Title: A stress response p38 MAP kinase inhibitor SB202190 promoted TFEB/TFE3-dependent autophagy and lysosomal biogenesis independent of p38

    Journal: Redox Biology

    doi: 10.1016/j.redox.2020.101445

    PPP3/calcineurin rather than MTOR signaling pathway is critical for SB202190-induced TFEB and TFE3 activation. ( A-B ) There were pronounced gel-shift of Flag-TFEB in CF-7 cells ( A ), or endogenous TFE3 in HeLa cells (B) in response to different concentrations of SB202190 (0–20 μM) or 20 μM of SB202190 for different durations (0–3 h), indicating that SB202190 caused TFEB and TFE3 dephosphorylation. ( C-D ) SB202190 did not inhibit MTOR signaling pathway. After cells were treated with indicated concentrations of SB202190 for 3 h, (at the time point that SB202190 robustly promotes TFEB/TFE3 to translocate into the nucleus) or 20 μM SB202190 for indicated durations, the expression of p-MTOR, MTOR, p-RPS6KB1/P70S6K, RPS6KB1/P70S6K, p-EIF4EBP1/p-4EBP1 and EIF4EBP1/4EBP1 were detected by Western blotting; Torin 1 (250 nm) was used as a positive control. ( E-F ) PPP3/calcineurin inhibitors FK506 plus CsA attenuated the translocation of TFEB from the cytoplasm into the nucleus in response to SB202190. After pretreatment of CF-7 cells with FK506 (10 μM) plus CsA (20 μM) for 30 min followed by adding SB202190 into cells for another 3 h, the cytoplasm and the nucleus distribution of TFEB were detected by immunostaining or Western blotting analysis. ( G ) FK506 plus CsA partially attenuated the gel-shift of Flag-TFEB in response to SB202190. After pretreatment of CF-7 cells with FK506 (10 μM) and CsA (20 μM) for 30 min followed by adding SB202190 (10 μM) into cells for another 3 h, the phosphorylation status of Flag-TFEB was determined. ( H–I ) FK506 and CsA attenuated the translocation of endogenous TFE3 from the cytoplasm into the nucleus in response to SB202190. After pretreatment of HeLa cells with FK506 (10 μM) plus CsA (20 μM) for 30 min followed by adding SB202190 into cells for another 3 h, the cytoplasm and the nucleus distribution of endogenous TFE3 were detected by immunostaining or Western blotting analysis. ( J ) FK506 plus CsA partially attenuated the gel-shift of endogenous TFE3 in response to SB202190. After pretreatment of HeLa cells with FK506 (10 μM) and CsA (20 μM) for 30 min followed by adding SB202190 (10 μM) into cells for another 3 h, the phosphorylation status of TFE3 was detected. Quantification of the number of cells with nuclear TFEB/TFE3 localization is presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group.
    Figure Legend Snippet: PPP3/calcineurin rather than MTOR signaling pathway is critical for SB202190-induced TFEB and TFE3 activation. ( A-B ) There were pronounced gel-shift of Flag-TFEB in CF-7 cells ( A ), or endogenous TFE3 in HeLa cells (B) in response to different concentrations of SB202190 (0–20 μM) or 20 μM of SB202190 for different durations (0–3 h), indicating that SB202190 caused TFEB and TFE3 dephosphorylation. ( C-D ) SB202190 did not inhibit MTOR signaling pathway. After cells were treated with indicated concentrations of SB202190 for 3 h, (at the time point that SB202190 robustly promotes TFEB/TFE3 to translocate into the nucleus) or 20 μM SB202190 for indicated durations, the expression of p-MTOR, MTOR, p-RPS6KB1/P70S6K, RPS6KB1/P70S6K, p-EIF4EBP1/p-4EBP1 and EIF4EBP1/4EBP1 were detected by Western blotting; Torin 1 (250 nm) was used as a positive control. ( E-F ) PPP3/calcineurin inhibitors FK506 plus CsA attenuated the translocation of TFEB from the cytoplasm into the nucleus in response to SB202190. After pretreatment of CF-7 cells with FK506 (10 μM) plus CsA (20 μM) for 30 min followed by adding SB202190 into cells for another 3 h, the cytoplasm and the nucleus distribution of TFEB were detected by immunostaining or Western blotting analysis. ( G ) FK506 plus CsA partially attenuated the gel-shift of Flag-TFEB in response to SB202190. After pretreatment of CF-7 cells with FK506 (10 μM) and CsA (20 μM) for 30 min followed by adding SB202190 (10 μM) into cells for another 3 h, the phosphorylation status of Flag-TFEB was determined. ( H–I ) FK506 and CsA attenuated the translocation of endogenous TFE3 from the cytoplasm into the nucleus in response to SB202190. After pretreatment of HeLa cells with FK506 (10 μM) plus CsA (20 μM) for 30 min followed by adding SB202190 into cells for another 3 h, the cytoplasm and the nucleus distribution of endogenous TFE3 were detected by immunostaining or Western blotting analysis. ( J ) FK506 plus CsA partially attenuated the gel-shift of endogenous TFE3 in response to SB202190. After pretreatment of HeLa cells with FK506 (10 μM) and CsA (20 μM) for 30 min followed by adding SB202190 (10 μM) into cells for another 3 h, the phosphorylation status of TFE3 was detected. Quantification of the number of cells with nuclear TFEB/TFE3 localization is presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group.

    Techniques Used: Activation Assay, Electrophoretic Mobility Shift Assay, De-Phosphorylation Assay, Expressing, Western Blot, Positive Control, Translocation Assay, Immunostaining

    Schematic illustration of the mechanism of action of SB202190 on activating TFEB/TFE3-mediated autophagy and lysosomal biogenesis. SB202190 causes the release of calcium from ER. Intracellular calcium subsequently activates the phosphatase PPP3/calcineurin, thereby de-phosphorylating and promoting TFEB and TFE3 nuclear accumulation. In the nucleus, TFEB and TFE3 transcriptionally regulate the expression of multiple autophagy- and lysosomal-related genes to enhance autophagy and lysosomal biogenesis. PPP3/calcineurin inhibitors FK506 and CsA, calcium chelator BAPTA-AM, or depleting ER calcium by TG (thapsigargin) all effectively attenuate TFEB and TFE3 activation in response to SB202190. Taken together, SB202190 activates TFEB- and TFE3-dependent autophagy and lysosomal biogenesis via ER calcium release and subsequent calcium-dependent PPP3/calcineurin activation, leading to dephosphorylation and activation of TFEB and TFE3.
    Figure Legend Snippet: Schematic illustration of the mechanism of action of SB202190 on activating TFEB/TFE3-mediated autophagy and lysosomal biogenesis. SB202190 causes the release of calcium from ER. Intracellular calcium subsequently activates the phosphatase PPP3/calcineurin, thereby de-phosphorylating and promoting TFEB and TFE3 nuclear accumulation. In the nucleus, TFEB and TFE3 transcriptionally regulate the expression of multiple autophagy- and lysosomal-related genes to enhance autophagy and lysosomal biogenesis. PPP3/calcineurin inhibitors FK506 and CsA, calcium chelator BAPTA-AM, or depleting ER calcium by TG (thapsigargin) all effectively attenuate TFEB and TFE3 activation in response to SB202190. Taken together, SB202190 activates TFEB- and TFE3-dependent autophagy and lysosomal biogenesis via ER calcium release and subsequent calcium-dependent PPP3/calcineurin activation, leading to dephosphorylation and activation of TFEB and TFE3.

    Techniques Used: Expressing, Activation Assay, De-Phosphorylation Assay

    Calcium is required for SB202190-induced TFEB and TFE3 activation as well as autophagy and lysosomal biogenesis. ( A ) SB202190 increased cytoplasmic Ca 2+ levels. Cytoplasmic Ca 2+ concentration in CF-7 cells was detected by using Fura-2 AM probe in Ca 2+ -free HBSS, followed by addition of indicated concentrations of SB202190. The peak value of 340/380 nm excitation ratio for fura-2 was quantified. At least 50 cells were recorded in each experiment and the experiments were repeated for 3 times. Data are presented as mean ± SEM of 3 replicates. ( B ) Calcium chelator BAPTA-AM reduced the translocation of TFEB from the cytoplasm into the nucleus in response to SB202190. After retreatment of CF-7 cells with BAPTA-AM (20 μM) for 30 min followed by being treated with SB202190 for another 3 h, the nucleus distribution of TFEB was detected by immunostaining. ( C ) Quantification of the number of cells with nuclear TFEB localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group. ( D ) Calcium chelator BAPTA-AM attenuated SB202190-induced accumulation of endogenous TFE3 in the nucleus. ( E ) Quantification of the number of cells with nuclear TFE3 localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. ( F-G ) Western blotting assay showed that calcium chelator BAPTA-AM reduces SB202190-induced translocation of TFEB and TFE3 from the cytoplasm into the nucleus. ( H ) Calcium chelator BAPTA-AM blocks SB202190-induced increase in lysosome contents as evidenced by Lysotracker Red staining. After pretreatment of HeLa cells with calcium chelator BAPTA-AM (20 μM) for 30 min followed by treatment with SB202190 for another 16 h, the lysosome contents were determined by flow cytometry after cells were loaded with LysoTracker Red DND-99 (75 nm, for 1 h). ( I ) Pretreatment of HeLa cell with BAPTA-AM reduced SB202190-inducedexpression of LAMP1, LC3B-II, and SQSTM1/p62 levels. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
    Figure Legend Snippet: Calcium is required for SB202190-induced TFEB and TFE3 activation as well as autophagy and lysosomal biogenesis. ( A ) SB202190 increased cytoplasmic Ca 2+ levels. Cytoplasmic Ca 2+ concentration in CF-7 cells was detected by using Fura-2 AM probe in Ca 2+ -free HBSS, followed by addition of indicated concentrations of SB202190. The peak value of 340/380 nm excitation ratio for fura-2 was quantified. At least 50 cells were recorded in each experiment and the experiments were repeated for 3 times. Data are presented as mean ± SEM of 3 replicates. ( B ) Calcium chelator BAPTA-AM reduced the translocation of TFEB from the cytoplasm into the nucleus in response to SB202190. After retreatment of CF-7 cells with BAPTA-AM (20 μM) for 30 min followed by being treated with SB202190 for another 3 h, the nucleus distribution of TFEB was detected by immunostaining. ( C ) Quantification of the number of cells with nuclear TFEB localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group. ( D ) Calcium chelator BAPTA-AM attenuated SB202190-induced accumulation of endogenous TFE3 in the nucleus. ( E ) Quantification of the number of cells with nuclear TFE3 localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. ( F-G ) Western blotting assay showed that calcium chelator BAPTA-AM reduces SB202190-induced translocation of TFEB and TFE3 from the cytoplasm into the nucleus. ( H ) Calcium chelator BAPTA-AM blocks SB202190-induced increase in lysosome contents as evidenced by Lysotracker Red staining. After pretreatment of HeLa cells with calcium chelator BAPTA-AM (20 μM) for 30 min followed by treatment with SB202190 for another 16 h, the lysosome contents were determined by flow cytometry after cells were loaded with LysoTracker Red DND-99 (75 nm, for 1 h). ( I ) Pretreatment of HeLa cell with BAPTA-AM reduced SB202190-inducedexpression of LAMP1, LC3B-II, and SQSTM1/p62 levels. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

    Techniques Used: Activation Assay, Concentration Assay, Translocation Assay, Immunostaining, Western Blot, Staining, Flow Cytometry

    ER calcium contributes to SB202190-induced TFEB and TFE3 activation as well as autophagy and lysosomal biogenesis. ( A ) Depleting ER Ca 2+ stores with thapsigargin (TG) reduced SB202190-induc ed increase of cytoplasmic calcium levels. In contrast, SB202190 still significantly increases cytoplasmic calcium levels after depleting lysosomal Ca 2+ stores with GPN (glycyl- l -phenylalanine-β-naphthylamide). The peak value of 340/380 nm excitation ratio for Fura-2 was quantified. At least 50 cells were recorded in each experiment and the experiments were repeated for 3 times. Data are presented as mean ± SEM of 3 replicates. ( B ) Depleting ER calcium stores with TG prevented the translocation of TFEB from the cytoplasm into the nucleus in response to SB202190. After pretreatment of CF-7 cells with TG (1 μM) for 30 min followed by being treated with SB202190 for another 3 h, the nucleus distribution of TFEB was detected by immunostaining. ( C ) Quantification of the number of cells with nuclear TFEB localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group. ( D ) TG attenuated SB202190-induced accumulation of endogenous TFE3 in the nucleus. ( E ) Quantification of the number of cells with nuclear TFE3 localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. ( F-G ) Effects of SB202190 on the expression of TFEB and endogenous TFE3 in the cytosolic (Cyt.) and nuclear (Nuc.) fractions were determined by Western blotting in the presence or the absence of TG, indicating that TG reduced SB202190-induced translocation of TFEB and TFE3 from the cytoplasm into the nucleus. (H ) TG reduces SB202190-induced increase of lysosome contents as evidenced by flow cytometry assay after cells were loaded with LysoTracker Red DND-99 (75 nm, for 1 h). ( I ) Pretreatment of HeLa cell with TG (1 μM) reduced SB202190-induced expression of LAMP1, LC3B-II, and SQSTM1/p62 levels. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
    Figure Legend Snippet: ER calcium contributes to SB202190-induced TFEB and TFE3 activation as well as autophagy and lysosomal biogenesis. ( A ) Depleting ER Ca 2+ stores with thapsigargin (TG) reduced SB202190-induc ed increase of cytoplasmic calcium levels. In contrast, SB202190 still significantly increases cytoplasmic calcium levels after depleting lysosomal Ca 2+ stores with GPN (glycyl- l -phenylalanine-β-naphthylamide). The peak value of 340/380 nm excitation ratio for Fura-2 was quantified. At least 50 cells were recorded in each experiment and the experiments were repeated for 3 times. Data are presented as mean ± SEM of 3 replicates. ( B ) Depleting ER calcium stores with TG prevented the translocation of TFEB from the cytoplasm into the nucleus in response to SB202190. After pretreatment of CF-7 cells with TG (1 μM) for 30 min followed by being treated with SB202190 for another 3 h, the nucleus distribution of TFEB was detected by immunostaining. ( C ) Quantification of the number of cells with nuclear TFEB localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group. ( D ) TG attenuated SB202190-induced accumulation of endogenous TFE3 in the nucleus. ( E ) Quantification of the number of cells with nuclear TFE3 localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. ( F-G ) Effects of SB202190 on the expression of TFEB and endogenous TFE3 in the cytosolic (Cyt.) and nuclear (Nuc.) fractions were determined by Western blotting in the presence or the absence of TG, indicating that TG reduced SB202190-induced translocation of TFEB and TFE3 from the cytoplasm into the nucleus. (H ) TG reduces SB202190-induced increase of lysosome contents as evidenced by flow cytometry assay after cells were loaded with LysoTracker Red DND-99 (75 nm, for 1 h). ( I ) Pretreatment of HeLa cell with TG (1 μM) reduced SB202190-induced expression of LAMP1, LC3B-II, and SQSTM1/p62 levels. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

    Techniques Used: Activation Assay, Translocation Assay, Immunostaining, Expressing, Western Blot, Flow Cytometry

    The p38 MAP kinase inhibitor SB202190 promoted the translocation of TFEB and TFE3 from the cytoplasm into the nucleus. ( A ) After being treated with SB202190 at indicated concentrations or a positive control Torin 1 (250 nm) for 3 h, HeLa cells stably expressing 3XFlag-TFEB (CF-7) were fixed and stained with the anti-Flag antibody (green) and DAPI (blue). Representative images are shown. ( B) Quantification of the number of cells with nuclear TFEB localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group. ( C ) The expression of Flag-TFEB in the cytosol (Cyt.) and the nucleus (Nuc.) were detected by Western blotting after being treated with indicated compounds for 3 h. ( D ) After being treated with SB202190 at indicated concentrations or a positive control Torin 1 (250 nm) for 3 h, cells were fixed and stained with anti-TFE3 antibody (green) and DAPI (blue). Representative images are shown. ( E ) Quantification of the number of cells with nuclear TFE3 localization. ( F ) The expression of endogenous TFE3 in the cytosol (Cyt.) and nucleus (Nuc.) were detected by Western blotting after being treated with indicated compounds for 3 h. ( G ) SB202190 promoted TFEB translocation from the cytoplasm into the nucleus in HEK293 cells. After transiently transfection of cells with Flag-TFEB plasmids for 48 h, followed by treating cells with SB202190 (20 μM) or a positive control Torin 1 (250 nm) for 3 h, cells were fixed and stained with the anti-Flag antibody (green) and DAPI (blue). Representative images are shown. ( H ) After being treated with SB202190 (10 μM) for 3 h, cells were fixed and stained with the TFE3 antibody (green) and DAPI (blue). Representative images are shown. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
    Figure Legend Snippet: The p38 MAP kinase inhibitor SB202190 promoted the translocation of TFEB and TFE3 from the cytoplasm into the nucleus. ( A ) After being treated with SB202190 at indicated concentrations or a positive control Torin 1 (250 nm) for 3 h, HeLa cells stably expressing 3XFlag-TFEB (CF-7) were fixed and stained with the anti-Flag antibody (green) and DAPI (blue). Representative images are shown. ( B) Quantification of the number of cells with nuclear TFEB localization. Data are presented as mean ± SEM of 3 replicates in a representative experiment. At least 200 cells were analyzed in each treatment group. ( C ) The expression of Flag-TFEB in the cytosol (Cyt.) and the nucleus (Nuc.) were detected by Western blotting after being treated with indicated compounds for 3 h. ( D ) After being treated with SB202190 at indicated concentrations or a positive control Torin 1 (250 nm) for 3 h, cells were fixed and stained with anti-TFE3 antibody (green) and DAPI (blue). Representative images are shown. ( E ) Quantification of the number of cells with nuclear TFE3 localization. ( F ) The expression of endogenous TFE3 in the cytosol (Cyt.) and nucleus (Nuc.) were detected by Western blotting after being treated with indicated compounds for 3 h. ( G ) SB202190 promoted TFEB translocation from the cytoplasm into the nucleus in HEK293 cells. After transiently transfection of cells with Flag-TFEB plasmids for 48 h, followed by treating cells with SB202190 (20 μM) or a positive control Torin 1 (250 nm) for 3 h, cells were fixed and stained with the anti-Flag antibody (green) and DAPI (blue). Representative images are shown. ( H ) After being treated with SB202190 (10 μM) for 3 h, cells were fixed and stained with the TFE3 antibody (green) and DAPI (blue). Representative images are shown. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

    Techniques Used: Translocation Assay, Positive Control, Stable Transfection, Expressing, Staining, Western Blot, Transfection

    SB202190 enhanced autophagy and lysosomal biogenesis. ( A ) SB202190 increased LC3B-II levels in a dose-dependent manner. After HeLa cells were treated with indicated concentrations of SB202190 or a positive control Torin 1 (250 nM) for 16 h, the expression of LC3B was detected by Western blotting. ( B ) HeLa cells were treated with SB202190 (10 μM) or a positive control Torin 1 (250 nM) for 16 h in the presence or absence of the lysosomal inhibitor chloroquine (CQ, 50 μM) (CQ was added into cells at last 3 h for drug treatment), then the expression of LC3B-II was detected and quantified. ( C ) HeLa cells were treated with SB202190 (10 μM) or a positive control Torin 1 (250 nM) for 16 h in the presence or absence of the v-ATPase inhibitor bafilomycin A1 to inhibit autophagy-lysosome fusion (Baf A1, 50 nM) (Baf A1 was added into cells at last 3 h for drug treatment). The expression of LC3B-II was detected and quantified. ( D ) After the treatment of HeLa cells stably expressing tf-LC3 plasmids with SB202190 (10 μM) for 16 h, the signal was captured by a fluorescence microscope and representative images are shown. ( E ) SB202190 increased LAMP1 levels in a dose-dependent manner. After HeLa cells were treated with indicated concentrations of SB202190 or a positive control, Torin 1 (250 nM) for 16 h, the expression of LAMP1 was detected by western blotting and quantified. ( F ) After HeLa cells were treated with SB202190 (10 μM) or Torin 1 (250 nM) for 16 h, cells were loaded with LysoTracker Red DND-99 (75 nm) for 1 h and the intensity was recorded by flow cytometer. Representative data are shown from three independent experiments. Quantitative data are presented as the mean ± SEM from at least 3 independent experiments. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
    Figure Legend Snippet: SB202190 enhanced autophagy and lysosomal biogenesis. ( A ) SB202190 increased LC3B-II levels in a dose-dependent manner. After HeLa cells were treated with indicated concentrations of SB202190 or a positive control Torin 1 (250 nM) for 16 h, the expression of LC3B was detected by Western blotting. ( B ) HeLa cells were treated with SB202190 (10 μM) or a positive control Torin 1 (250 nM) for 16 h in the presence or absence of the lysosomal inhibitor chloroquine (CQ, 50 μM) (CQ was added into cells at last 3 h for drug treatment), then the expression of LC3B-II was detected and quantified. ( C ) HeLa cells were treated with SB202190 (10 μM) or a positive control Torin 1 (250 nM) for 16 h in the presence or absence of the v-ATPase inhibitor bafilomycin A1 to inhibit autophagy-lysosome fusion (Baf A1, 50 nM) (Baf A1 was added into cells at last 3 h for drug treatment). The expression of LC3B-II was detected and quantified. ( D ) After the treatment of HeLa cells stably expressing tf-LC3 plasmids with SB202190 (10 μM) for 16 h, the signal was captured by a fluorescence microscope and representative images are shown. ( E ) SB202190 increased LAMP1 levels in a dose-dependent manner. After HeLa cells were treated with indicated concentrations of SB202190 or a positive control, Torin 1 (250 nM) for 16 h, the expression of LAMP1 was detected by western blotting and quantified. ( F ) After HeLa cells were treated with SB202190 (10 μM) or Torin 1 (250 nM) for 16 h, cells were loaded with LysoTracker Red DND-99 (75 nm) for 1 h and the intensity was recorded by flow cytometer. Representative data are shown from three independent experiments. Quantitative data are presented as the mean ± SEM from at least 3 independent experiments. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

    Techniques Used: Positive Control, Expressing, Western Blot, Stable Transfection, Fluorescence, Microscopy, Flow Cytometry

    SB202190-induced TFEB and TFE3 nuclear accumulation is independent of p38 MAP kinase inhibition. ( A ) After CF-7 cells were treated with p38 MAP kinase inhibitors SB202190, BIRB-796, SB203580, and SB239063 at the concentration of 10 μM for 3 h, cellular distribution of TFEB was detected by immunostaining. ( B ) After HeLa cells were treated with p38 MAP kinase inhibitors SB202190, BIRB796, SB203580, and SB239063 at the concentration of 10 μM for 3 h, cellular distribution of endogenous TFE3 was detected by immunostaining. ( C ) Western blotting results showed that SB202190, rather than other p38 MAP kinase inhibitors (10 μM for 16 h) robustly increases LC3B-II and SQSTM1/p62 levels in HeLa cells. ( D ) SB202190 still increased the expression of LAMP1 and LC3B-II levels after knockdown of Mapk14/p38 α. ( E -F ) After HeLa were transfected with non-target of MAPK14/p38α specific siRNA for 48 h, followed by treatment with SB202190 for 3 h, the cellular distribution of TFEB (E) and TFE3 (F) were detected by immunostaining assay.
    Figure Legend Snippet: SB202190-induced TFEB and TFE3 nuclear accumulation is independent of p38 MAP kinase inhibition. ( A ) After CF-7 cells were treated with p38 MAP kinase inhibitors SB202190, BIRB-796, SB203580, and SB239063 at the concentration of 10 μM for 3 h, cellular distribution of TFEB was detected by immunostaining. ( B ) After HeLa cells were treated with p38 MAP kinase inhibitors SB202190, BIRB796, SB203580, and SB239063 at the concentration of 10 μM for 3 h, cellular distribution of endogenous TFE3 was detected by immunostaining. ( C ) Western blotting results showed that SB202190, rather than other p38 MAP kinase inhibitors (10 μM for 16 h) robustly increases LC3B-II and SQSTM1/p62 levels in HeLa cells. ( D ) SB202190 still increased the expression of LAMP1 and LC3B-II levels after knockdown of Mapk14/p38 α. ( E -F ) After HeLa were transfected with non-target of MAPK14/p38α specific siRNA for 48 h, followed by treatment with SB202190 for 3 h, the cellular distribution of TFEB (E) and TFE3 (F) were detected by immunostaining assay.

    Techniques Used: Inhibition, Concentration Assay, Immunostaining, Western Blot, Expressing, Transfection

    TFEB and TFE3 are necessary for SB202190-induced autophagy and lysosomal biogenesis. ( A ) SB202190 increased the expression of multiple autophagy-lysosomal pathway related genes. After HeLa cells were treated with SB202190 (10 μM) for 12 h, the expression of several autophagy- and lysosome-related genes were detected by real-time PCR. ( B ) Quantification data of TFEB and TFE3 levels after transfection of HeLa cells with TFEB and TFE3 siRNAs for 48 h. ( C ) Knockdown of the expression of Tfeb and Tfe3 attenuated SB202190-induced expression of several autophagy and lysosome-related genes as reflected by real-time PCR assay. ( D ) Tfeb or/and Tfe3 knockdown attenuated SB202190-induced expression of LAMP1, SQSTM1/p62 and LC3B-II levels in HeLa cells. HeLa cells were treated with SB202190 (10 μM) for 16 h after transfection of cells with indicated SiRNAs for 48 h. The expressions of LAMP1, SQSTM1/p62, MAP1LC3, TFEB, and TFE3 were detected. ( E ) Quantification data showed that depletion of TFEB/TFE3 partially but significantly attenuated SB202190-induced expression of LC3B-II levels. Quantitative data are presented as the mean ± SEM from at least 3 independent experiments.
    Figure Legend Snippet: TFEB and TFE3 are necessary for SB202190-induced autophagy and lysosomal biogenesis. ( A ) SB202190 increased the expression of multiple autophagy-lysosomal pathway related genes. After HeLa cells were treated with SB202190 (10 μM) for 12 h, the expression of several autophagy- and lysosome-related genes were detected by real-time PCR. ( B ) Quantification data of TFEB and TFE3 levels after transfection of HeLa cells with TFEB and TFE3 siRNAs for 48 h. ( C ) Knockdown of the expression of Tfeb and Tfe3 attenuated SB202190-induced expression of several autophagy and lysosome-related genes as reflected by real-time PCR assay. ( D ) Tfeb or/and Tfe3 knockdown attenuated SB202190-induced expression of LAMP1, SQSTM1/p62 and LC3B-II levels in HeLa cells. HeLa cells were treated with SB202190 (10 μM) for 16 h after transfection of cells with indicated SiRNAs for 48 h. The expressions of LAMP1, SQSTM1/p62, MAP1LC3, TFEB, and TFE3 were detected. ( E ) Quantification data showed that depletion of TFEB/TFE3 partially but significantly attenuated SB202190-induced expression of LC3B-II levels. Quantitative data are presented as the mean ± SEM from at least 3 independent experiments.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Transfection

    8) Product Images from "Wnt/β-catenin signaling pathway induces autophagy-mediated temozolomide-resistance in human glioblastoma"

    Article Title: Wnt/β-catenin signaling pathway induces autophagy-mediated temozolomide-resistance in human glioblastoma

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-020-02988-8

    ATG9B expression was regulated through Wnt/β-catenin signaling pathway. a , b The mRNA expression of ATG9B detected by real-time PCR. DAB2IP-low cells were co-treated with 50µM TMZ and small molecule inhibitors (LGK974, 500nM; LY3214996, 50nM; Rapamycin, 10nM; LY294002 1µM; and SB202190, 10µM) for 48h before the analysis ( a ). DAB2IP-high cells were co-treated with 50µM TMZ and Wnt signaling activator LiCl for 48h before the analysis ( b ). c TMZ log–dose–response analysis (IC 50 ). Cells were seeded in a 96-well and treated with increment doses of TMZ as indicated for 48h. LGK974 (200nM) was treated in combination with TMZ as indicated. d Colony formation assay. DAB2IP-low A172 KD and LN229 Vc cells were seeded in 6-well plates at a density of 1000 cells per well. TMZ, LGK974, and a combination of TMZ and LGK974 were treated to cells for 10 days. e Cell viability tested by MTT assay. Cells were treated with 50µM TMZ, 200nM LGK974, or a combination for 48h. Percent cell survival was normalized to untreated conditions. Drug synergistic effects were determined based on the combination index (CI). CI
    Figure Legend Snippet: ATG9B expression was regulated through Wnt/β-catenin signaling pathway. a , b The mRNA expression of ATG9B detected by real-time PCR. DAB2IP-low cells were co-treated with 50µM TMZ and small molecule inhibitors (LGK974, 500nM; LY3214996, 50nM; Rapamycin, 10nM; LY294002 1µM; and SB202190, 10µM) for 48h before the analysis ( a ). DAB2IP-high cells were co-treated with 50µM TMZ and Wnt signaling activator LiCl for 48h before the analysis ( b ). c TMZ log–dose–response analysis (IC 50 ). Cells were seeded in a 96-well and treated with increment doses of TMZ as indicated for 48h. LGK974 (200nM) was treated in combination with TMZ as indicated. d Colony formation assay. DAB2IP-low A172 KD and LN229 Vc cells were seeded in 6-well plates at a density of 1000 cells per well. TMZ, LGK974, and a combination of TMZ and LGK974 were treated to cells for 10 days. e Cell viability tested by MTT assay. Cells were treated with 50µM TMZ, 200nM LGK974, or a combination for 48h. Percent cell survival was normalized to untreated conditions. Drug synergistic effects were determined based on the combination index (CI). CI

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Colony Assay, MTT Assay

    9) Product Images from "A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway"

    Article Title: A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway

    Journal: Journal of Developmental Biology

    doi: 10.3390/jdb8030012

    p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.
    Figure Legend Snippet: p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.

    Techniques Used: Concentration Assay

    Related Articles

    Recombinant:

    Article Title: Pyrimethamine Elicits Antitumor Effects on Prostate Cancer by Inhibiting the p38-NF-κB Pathway
    Article Snippet: ReagentsPyrimethamine (purchased from Sigma, ShangHai, China) was dissolved in dimethyl sulfoxide (DMSO) to a final concentration of 100 mmol/L. .. The p38 MAPK inhibitor, SB202190 (FHPI), was procured from Selleck Chemicals (Houston, USA), and the recombinant tumor necrosis factor alpha (TNF-α) was obtained from Huaxia Ocean Technology (Beijing, China). .. In all the experiments, the final DMSO concentration was < 0.1%, and DMSO alone had no noticeable effects on the cultured cells.

    other:

    Article Title: A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway
    Article Snippet: After 18–20 h of starvation, the cells were treated with the MEK inhibitor U0126-EtOH (U0126) at 1 µM, NF-κB inhibitor caffeic acid phenethyl ester (CAPE) at 1 µM, or p38 inhibitor SB202190 at 10 µM or left untreated.

    Article Title: A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway
    Article Snippet: The cells were treated as follows: RANKL, RANKL + inhibitor (U0126, CAPE, or SB202190), RANKL + CK2.3, RANKL + CK2.3 + inhibitor (U0126, CAPE, or SB202190), RANKL + BMP2, RANKL + BMP2 + inhibitor (U0126, CAPE, or SB202190), untreated, and untreated + inhibitor (U0126, CAPE, or SB202190).

    Article Title: The Esophageal Organoid System Reveals Functional Interplay Between Notch and Cytokines in Reactive Epithelial Changes
    Article Snippet: Among factors and agents originally used for murine esophageal 3D organoids, R-Spondin1 and Noggin, Wnt3A, or A83-01 improved OFR in passaged organoids in KSFMC; however, SB202190, a p38 mitogen-activated protein kinase inhibitor, impaired organoid formation by human cells.

    Article Title: Bafilomycin A1 induces caspase-independent cell death in hepatocellular carcinoma cells via targeting of autophagy and MAPK pathways
    Article Snippet: The inhibitors PD98059, SB202190, SP600125 and AZ191 were purchased from Selleck.

    Cell Culture:

    Article Title: Protective mechanism of FSH against oxidative damage in mouse ovarian granulosa cells by repressing autophagy
    Article Snippet: The cells were cultured in DMEM/F-12 (1:1) medium (Life Technologies, 11330–057) supplemented with 10% fetal bovine serum (Gibco, 10270) and 100 units/ml penicillin plus 100 μg/ml streptomycin (Gibco, 15140) for 4 d at 37°C with 5% CO2 . .. For FSH treatment, GCs exposed to 200 μM H2 O2 (Sigma-Aldrich, 216763–100ML) for 0 h (none) or 1 h were washed in PBS, and cultured with serum-free DMEM/F-12 containing 7.5 IU/ml FSH (Sigma-Aldrich, F4021) in the presence or absence of pepstatin A and E64 (10 μg/ml; Selleck, S7381 and S7379, respectively), perifosine (10 μM; Selleck, S1037), SB203580 (20 μM; Selleck, S1077), U0126 (3 μM; Selleck, S1102), torin 1 (100 nM; Selleck, S2827), sirtinol (100 μM; Selleck, S2804) or SRT1720 (100 μM; Selleck, S1129) for 0.5, 1, 2, or 3 h as indicated. .. In some experiments, GCs were treated with chloroquine (50 μM; Sigma-Aldrich, C6628) or Z-VAD-FMK (50 μM; Selleck, S7023) for 0.5, 1, 2, or 3 h following 1 h of H2 O2 incubation.

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    Selleck Chemicals sb202190
    Optimization of normal human and murine esophageal 3D organoid culture . Esophageal 3D organoid culture was optimized with the normal human esophageal epithelial cell line EPC2-hTERT in A–C and wild-type murine esophageal cells dissociated from epithelial sheets in D–F (ie, Noggin/R-Spondin1-conditioned medium, Wnt3A, A83-01, <t>SB202190,</t> gastrin, and nicotinamide) were added as indicated. KSFM contains 0.09 mM Ca 2+ (low Ca 2+ ), whereas KSFMC is supplemented with additional CaCl 2 to the final concentration of 0.6 mM Ca 2+ (high Ca 2+ ). The aDMEM medium contains 1 mM Ca 2+ . 3D culture products were photomicrographed under a phase-contrast microscopy and further recovered from Matrigel for H E staining in A and D . Organoids were defined as 3D structures with a diameter of 50 μm or larger. Note that 3D organoid formation was barely observed in aDMEM/F12 with or without unique supplements. Addition of unique supplements also limited organoid formation in KSFM. 3D organoids grown in KSFM showed a lobular pattern of basaloid cell expansion toward surrounding matrix and had a less explicit differentiation gradient as compared with 3D organoids formed in KSFMC. Average organoid size was determined in B and E . OFR was determined in C and F . Scale bar = 100 μm in phase-contrast images and 50 μm in H E staining. * P
    Sb202190, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sb202190/product/Selleck Chemicals
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    sb202190 - by Bioz Stars, 2021-03
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    Selleck Chemicals sb202190 fhpi
    p38 inhibitor, <t>SB202190,</t> did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.
    Sb202190 Fhpi, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sb202190 fhpi/product/Selleck Chemicals
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sb202190 fhpi - by Bioz Stars, 2021-03
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    Optimization of normal human and murine esophageal 3D organoid culture . Esophageal 3D organoid culture was optimized with the normal human esophageal epithelial cell line EPC2-hTERT in A–C and wild-type murine esophageal cells dissociated from epithelial sheets in D–F (ie, Noggin/R-Spondin1-conditioned medium, Wnt3A, A83-01, SB202190, gastrin, and nicotinamide) were added as indicated. KSFM contains 0.09 mM Ca 2+ (low Ca 2+ ), whereas KSFMC is supplemented with additional CaCl 2 to the final concentration of 0.6 mM Ca 2+ (high Ca 2+ ). The aDMEM medium contains 1 mM Ca 2+ . 3D culture products were photomicrographed under a phase-contrast microscopy and further recovered from Matrigel for H E staining in A and D . Organoids were defined as 3D structures with a diameter of 50 μm or larger. Note that 3D organoid formation was barely observed in aDMEM/F12 with or without unique supplements. Addition of unique supplements also limited organoid formation in KSFM. 3D organoids grown in KSFM showed a lobular pattern of basaloid cell expansion toward surrounding matrix and had a less explicit differentiation gradient as compared with 3D organoids formed in KSFMC. Average organoid size was determined in B and E . OFR was determined in C and F . Scale bar = 100 μm in phase-contrast images and 50 μm in H E staining. * P

    Journal: Cellular and Molecular Gastroenterology and Hepatology

    Article Title: The Esophageal Organoid System Reveals Functional Interplay Between Notch and Cytokines in Reactive Epithelial Changes

    doi: 10.1016/j.jcmgh.2017.12.013

    Figure Lengend Snippet: Optimization of normal human and murine esophageal 3D organoid culture . Esophageal 3D organoid culture was optimized with the normal human esophageal epithelial cell line EPC2-hTERT in A–C and wild-type murine esophageal cells dissociated from epithelial sheets in D–F (ie, Noggin/R-Spondin1-conditioned medium, Wnt3A, A83-01, SB202190, gastrin, and nicotinamide) were added as indicated. KSFM contains 0.09 mM Ca 2+ (low Ca 2+ ), whereas KSFMC is supplemented with additional CaCl 2 to the final concentration of 0.6 mM Ca 2+ (high Ca 2+ ). The aDMEM medium contains 1 mM Ca 2+ . 3D culture products were photomicrographed under a phase-contrast microscopy and further recovered from Matrigel for H E staining in A and D . Organoids were defined as 3D structures with a diameter of 50 μm or larger. Note that 3D organoid formation was barely observed in aDMEM/F12 with or without unique supplements. Addition of unique supplements also limited organoid formation in KSFM. 3D organoids grown in KSFM showed a lobular pattern of basaloid cell expansion toward surrounding matrix and had a less explicit differentiation gradient as compared with 3D organoids formed in KSFMC. Average organoid size was determined in B and E . OFR was determined in C and F . Scale bar = 100 μm in phase-contrast images and 50 μm in H E staining. * P

    Article Snippet: Given the inhibitory effect of SB202190, human esophageal cells may be sensitive to p38 mitogen-activated protein kinase–mediated processes, raising the possibility that antioxidants may alleviate cellular stress and improve cell survival.

    Techniques: Concentration Assay, Microscopy, Staining

    p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.

    Journal: Journal of Developmental Biology

    Article Title: A Synthetic Peptide, CK2.3, Inhibits RANKL-Induced Osteoclastogenesis through BMPRIa and ERK Signaling Pathway

    doi: 10.3390/jdb8030012

    Figure Lengend Snippet: p38 inhibitor, SB202190, did not affect the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. ( A ) Concentration curve for SB202190 to determine the working concentration. ( B ) The effect of SB202190 on the antagonistic effect of CK2.3 on RANKL-induced osteoclastogenesis. a: denotes significant difference from Control without inhibitor, b: denotes significant difference from Control with inhibitor, c: denotes significant difference from RANKL at 10 ng/mL without inhibitor, d: denotes significant difference from RANKL at 10 ng/mL with inhibitor, and e: denotes significant difference from RANKL + CK2.3 at 100 nM without inhibitor.

    Article Snippet: The cells were treated as follows: RANKL, RANKL + inhibitor (U0126, CAPE, or SB202190), RANKL + CK2.3, RANKL + CK2.3 + inhibitor (U0126, CAPE, or SB202190), RANKL + BMP2, RANKL + BMP2 + inhibitor (U0126, CAPE, or SB202190), untreated, and untreated + inhibitor (U0126, CAPE, or SB202190).

    Techniques: Concentration Assay

    Pyrimethamine inhibits proliferation and promotes apoptosis in CRPC cells through the inhibition of p38 MAPK. (A, B) DU145 and PC3 cells were treated with pyrimethamine or SB202190 (10 μM) for 24 h and then incubated with TNF-a (10 ng/ml) for 24 h, and quantification of percentages in cell cycle phases was analyzed by flow cytometry. (C, D) Mean percentage of cells ± SD was shown (DU145 and PC3). **:P

    Journal: Frontiers in Pharmacology

    Article Title: Pyrimethamine Elicits Antitumor Effects on Prostate Cancer by Inhibiting the p38-NF-κB Pathway

    doi: 10.3389/fphar.2020.00758

    Figure Lengend Snippet: Pyrimethamine inhibits proliferation and promotes apoptosis in CRPC cells through the inhibition of p38 MAPK. (A, B) DU145 and PC3 cells were treated with pyrimethamine or SB202190 (10 μM) for 24 h and then incubated with TNF-a (10 ng/ml) for 24 h, and quantification of percentages in cell cycle phases was analyzed by flow cytometry. (C, D) Mean percentage of cells ± SD was shown (DU145 and PC3). **:P

    Article Snippet: The p38 MAPK inhibitor, SB202190 (FHPI), was procured from Selleck Chemicals (Houston, USA), and the recombinant tumor necrosis factor alpha (TNF-α) was obtained from Huaxia Ocean Technology (Beijing, China).

    Techniques: Inhibition, Incubation, Flow Cytometry

    Pyrimethamine regulates the p65/NF- k B pathway in CRPC cells through p38 MAPK inhibition. (A, B) The western blot analysis of the expression of p38, pp38, and pp65 after treatment of 0, 32, and 100 μM pyrimethamine for 24 h in DU145 and PC3 cell lines. The ratio of pp38/p38 was normalized to GAPDH. (C, D) DU145 and PC3 cells were treated for 24 h with pyrimethamine or SB202190 (10 μM) and then treated for 6 h in the absence or presence of TNF-a (10 ng/ml). Total cell lysates were analyzed by western blot with antibodies against p38, pp38, and pP65. (E) A consolidated model that illustrates a plausible sequence for the mechanism by which pyrimethamine elicits antitumor effects in prostate cancer.

    Journal: Frontiers in Pharmacology

    Article Title: Pyrimethamine Elicits Antitumor Effects on Prostate Cancer by Inhibiting the p38-NF-κB Pathway

    doi: 10.3389/fphar.2020.00758

    Figure Lengend Snippet: Pyrimethamine regulates the p65/NF- k B pathway in CRPC cells through p38 MAPK inhibition. (A, B) The western blot analysis of the expression of p38, pp38, and pp65 after treatment of 0, 32, and 100 μM pyrimethamine for 24 h in DU145 and PC3 cell lines. The ratio of pp38/p38 was normalized to GAPDH. (C, D) DU145 and PC3 cells were treated for 24 h with pyrimethamine or SB202190 (10 μM) and then treated for 6 h in the absence or presence of TNF-a (10 ng/ml). Total cell lysates were analyzed by western blot with antibodies against p38, pp38, and pP65. (E) A consolidated model that illustrates a plausible sequence for the mechanism by which pyrimethamine elicits antitumor effects in prostate cancer.

    Article Snippet: The p38 MAPK inhibitor, SB202190 (FHPI), was procured from Selleck Chemicals (Houston, USA), and the recombinant tumor necrosis factor alpha (TNF-α) was obtained from Huaxia Ocean Technology (Beijing, China).

    Techniques: Inhibition, Western Blot, Expressing, Sequencing