bix 02189  (MedChemExpress)

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: The MEK5/ERK5 pathway promotes the activation of the Hedgehog/GLI signaling in melanoma cells

doi: 10.1007/s13402-025-01050-z

Figure Lengend Snippet: ERK5 activation increases GLI1/GLI2 protein levels and transcriptional activity in NIH/3T3 cells. ( A ) Cells transduced with lentiviral vectors carrying control non-targeting shRNA (shNT) or murine ERK5-specific shRNA (shERK5-1) were transfected with pcDNA3.1 (-) or pCMV5-MEK5DD-HA and with GLI reporter (GLI-BS) for 12 h, before being treated with vehicle (-) or SAG for 48 h. Relative luciferase activity (RLU) was firefly/Renilla ratios normalized for control ± SD ( n = 3). ( B - C ) Cells transfected with pcDNA3.1 or pCMV5-MEK5DD-HA were treated with vehicle (-) or SAG for 48 h. Western Blot was performed. Graphs show averages of relative integrated density (RID) ± SD ( n = 3). ( D ) 24-hour serum-deprived cells were treated with SAG and/or BIX02189. Western Blot was performed. Graphs show averages of RID ± SD ( n = 3)

Article Snippet: ERK5 inhibitors JWG-071 [ ], AX15836 [ ] and XMD8-92 [ ], MEK5 inhibitors GW284543 [ ] and BIX02189 [ ] (MedChemExpress LLC, Princeton, NJ, U.S.A.), the GLI1/2 inhibitor GANT61 [ ] and the SMO agonist SAG [ ] (Sigma-Aldrich, St Louis, MO, U.S.A. and MedChemExpress LLC, Princeton, NJ, U.S.A.) were dissolved in DMSO.

Techniques: Activation Assay, Activity Assay, Transduction, Control, shRNA, Transfection, Luciferase, Western Blot

Journal: Cellular Oncology (Dordrecht, Netherlands)

Article Title: The MEK5/ERK5 pathway promotes the activation of the Hedgehog/GLI signaling in melanoma cells

doi: 10.1007/s13402-025-01050-z

Figure Lengend Snippet: Combined targeting of MEK5 and GLI synergistically reduces the volume of melanoma spheroids. ( A - B ) A375 ( A ) and SSM2c ( B ) spheroids were treated with DMSO (-), GANT-61, GW284543 or BIX02189 or with the combinations at the indicated concentrations. Graphs show the quantification of spheroid volume after 5 days normalized for the time point 0. Data represent mean ± SD from three independent experiments. Representative images of spheroids taken at day 5 are shown. § Bliss independence score (> 0) indicates synergistic effects over single treatments. Bliss score = 0.027 (GANT + GW284543 ) or 0.008 (GANT + BIX02189) in A375 cells; Bliss score = 0.291 (GANT + GW284543 ) or 0.327 (GANT + BIX02189) in SSM2c cells

Article Snippet: ERK5 inhibitors JWG-071 [ ], AX15836 [ ] and XMD8-92 [ ], MEK5 inhibitors GW284543 [ ] and BIX02189 [ ] (MedChemExpress LLC, Princeton, NJ, U.S.A.), the GLI1/2 inhibitor GANT61 [ ] and the SMO agonist SAG [ ] (Sigma-Aldrich, St Louis, MO, U.S.A. and MedChemExpress LLC, Princeton, NJ, U.S.A.) were dissolved in DMSO.

Techniques:

Journal: FEBS Open Bio

Article Title: Ulinastatin promotes macrophage efferocytosis and ameliorates lung inflammation via the ERK5 /Mer signaling pathway

doi: 10.1002/2211-5463.13461

Figure Lengend Snippet: Ulinastatin upregulated Mer expression by activating ERK5. (A) ERK5 and p‐ERK5 protein levels were detected by Western blotting. Upper part: Representative blot of protein expression; lower part: Relative protein expression of ERK5 and p‐ERK5. (B) ERK5 and p‐ERK5 protein levels were determined by Western blotting after treatment with inhibitor BIX02189. Upper part: Representative blot of protein expression; lower part: Relative protein expression of ERK5 and p‐ERK5. (C) ERK5 protein levels were determined by Western blotting after treatment with the inhibitor XMD8‐92. Left: Representative blot of protein expression; right: Relative protein expression of ERK5 and p‐ERK5. (D) Immunofluorescence assay for the determination of membrane protein Mer after treatment with the inhibitor BIX02189. (E) Relative protein expression of membrane protein Mer. (F) The level of membrane protein Mer was determined by immunofluorescence assay after treatment with the inhibitor XMD8‐92. (G) Relative protein expression of membrane protein Mer. Data is expressed as mean ± standard deviation, one‐way ANOVA, n = 3, * P < 0.05 vs. 0 U·mL −1 UTI group. [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: After cell adherence, the cells were pretreated with ulinastatin at different concentrations for 3 h. ERK5 inhibitor BIX02189 (MedChemExpress, HY‐12056) was used to divide the cells into blank control group, BIX02189 group, 5000 U·mL −1 group, and 5000 U·mL −1 + BIX02189 group.

Techniques: Expressing, Western Blot, Immunofluorescence, Membrane, Standard Deviation

Journal: FEBS Open Bio

Article Title: Ulinastatin promotes macrophage efferocytosis and ameliorates lung inflammation via the ERK5 /Mer signaling pathway

doi: 10.1002/2211-5463.13461

Figure Lengend Snippet: Ulinastatin enhanced macrophage efferocytosis and ameliorated LPS‐induced inflammatory injury. (A) Effect of UTI on phagocytosis and apoptosis of HL60 cells by RAW264.7 cells after using BIX02189. Left panel shows representative fluorescence images of HL60 phagocytic apoptosis in RAW264.7 cells. Scale, 100 μm; (B) UTI enhanced the ability of alveolar macrophage to phagocytose apoptotic PMNs after use of BIX02189. Representative images of apoptotic PMNs from alveolar macrophage phagocytizing cells, scale:100 μm; arrows indicate macrophages containing apoptotic bodies. (C) Hematoxylin and eosin (H&E) staining of mouse lung tissue after use of BIX02189, scale: 200 μm; (D) UTI enhanced the ability of alveolar macrophage to phagocytose apoptotic PMNs with XMD8‐92. Representative images of apoptotic PMNs from alveolar macrophages phagocytizing cells, scale:100 μm; arrows indicate macrophages containing apoptotic bodies. (E) Hematoxylin and eosin (H&E) staining of mouse lung tissue after use of XMD8‐92, scale: 200 μm; (F) Phagocytosis index (BIX02189) based on fluorescence images. (G) Phagocytosis index (BIX02189) based on representative images. (H) H&E staining was used to assess the pathological score (BIX02189). (I) Phagocytosis index (XMD8‐92) based on representative images. (J) H&E staining was used to assess the pathological score (XMD8‐92). Data are presented as mean ± standard deviation, one‐way ANOVA, n = 3. * P < 0.05 vs. UTI + LPS group or 5000 U·mL −1 UTI group. [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: After cell adherence, the cells were pretreated with ulinastatin at different concentrations for 3 h. ERK5 inhibitor BIX02189 (MedChemExpress, HY‐12056) was used to divide the cells into blank control group, BIX02189 group, 5000 U·mL −1 group, and 5000 U·mL −1 + BIX02189 group.

Techniques: Fluorescence, Staining, Standard Deviation

Journal: FEBS Open Bio

Article Title: Ulinastatin promotes macrophage efferocytosis and ameliorates lung inflammation via the ERK5 /Mer signaling pathway

doi: 10.1002/2211-5463.13461

Figure Lengend Snippet: Ulinastatin upregulated Mer expression by activating ERK5. (A) ERK5 and p‐ERK5 protein levels were detected by Western blotting. Upper part: Representative blot of protein expression; lower part: Relative protein expression of ERK5 and p‐ERK5. (B) ERK5 and p‐ERK5 protein levels were determined by Western blotting after treatment with inhibitor BIX02189. Upper part: Representative blot of protein expression; lower part: Relative protein expression of ERK5 and p‐ERK5. (C) ERK5 protein levels were determined by Western blotting after treatment with the inhibitor XMD8‐92. Left: Representative blot of protein expression; right: Relative protein expression of ERK5 and p‐ERK5. (D) Immunofluorescence assay for the determination of membrane protein Mer after treatment with the inhibitor BIX02189. (E) Relative protein expression of membrane protein Mer. (F) The level of membrane protein Mer was determined by immunofluorescence assay after treatment with the inhibitor XMD8‐92. (G) Relative protein expression of membrane protein Mer. Data is expressed as mean ± standard deviation, one‐way ANOVA, n = 3, * P < 0.05 vs. 0 U·mL −1 UTI group. [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: After cell adherence, the cells were pretreated with ulinastatin at different concentrations for 3 h. ERK5 inhibitor BIX02189 (MedChemExpress, HY‐12056) was used to divide the cells into blank control group, BIX02189 group, 5000 U·mL −1 group, and 5000 U·mL −1 + BIX02189 group.

Techniques: Expressing, Western Blot, Immunofluorescence, Membrane, Standard Deviation

Journal: FEBS Open Bio

Article Title: Ulinastatin promotes macrophage efferocytosis and ameliorates lung inflammation via the ERK5 /Mer signaling pathway

doi: 10.1002/2211-5463.13461

Figure Lengend Snippet: Ulinastatin enhanced macrophage efferocytosis and ameliorated LPS‐induced inflammatory injury. (A) Effect of UTI on phagocytosis and apoptosis of HL60 cells by RAW264.7 cells after using BIX02189. Left panel shows representative fluorescence images of HL60 phagocytic apoptosis in RAW264.7 cells. Scale, 100 μm; (B) UTI enhanced the ability of alveolar macrophage to phagocytose apoptotic PMNs after use of BIX02189. Representative images of apoptotic PMNs from alveolar macrophage phagocytizing cells, scale:100 μm; arrows indicate macrophages containing apoptotic bodies. (C) Hematoxylin and eosin (H&E) staining of mouse lung tissue after use of BIX02189, scale: 200 μm; (D) UTI enhanced the ability of alveolar macrophage to phagocytose apoptotic PMNs with XMD8‐92. Representative images of apoptotic PMNs from alveolar macrophages phagocytizing cells, scale:100 μm; arrows indicate macrophages containing apoptotic bodies. (E) Hematoxylin and eosin (H&E) staining of mouse lung tissue after use of XMD8‐92, scale: 200 μm; (F) Phagocytosis index (BIX02189) based on fluorescence images. (G) Phagocytosis index (BIX02189) based on representative images. (H) H&E staining was used to assess the pathological score (BIX02189). (I) Phagocytosis index (XMD8‐92) based on representative images. (J) H&E staining was used to assess the pathological score (XMD8‐92). Data are presented as mean ± standard deviation, one‐way ANOVA, n = 3. * P < 0.05 vs. UTI + LPS group or 5000 U·mL −1 UTI group. [Colour figure can be viewed at wileyonlinelibrary.com ]

Article Snippet: After cell adherence, the cells were pretreated with ulinastatin at different concentrations for 3 h. ERK5 inhibitor BIX02189 (MedChemExpress, HY‐12056) was used to divide the cells into blank control group, BIX02189 group, 5000 U·mL −1 group, and 5000 U·mL −1 + BIX02189 group.

Techniques: Fluorescence, Staining, Standard Deviation