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: Pathogens

Article Title: Purinergic Receptor Antagonists Inhibit Hemolysis Induced by Clostridium perfringens Alpha Toxin

doi: 10.3390/pathogens13060454

Figure Lengend Snippet: Effects of different pathway inhibitors on the rCPA-induced hemolysis of human and murine erythrocytes. ( a ) The degree of hemolysis did not change with increasing concentrations of the p38 MAPK inhibitor BIRB 796, implying no involvement in rCPA-induced hemolysis, in contrast to ( b ) the Akt inhibitor API-2, which can significantly inhibit rCPA-induced hemolysis. Regarding the MEK/ERK pathway, ( c ) the ERK inhibitor FR180204 slightly inhibits rCPA-induced hemolysis, ( d ) the MEK1 inhibitor PD334581 and ( e ) MEK1/2 inhibitor SL327 significantly inhibit rCPA-induced hemolysis, in contrast to ( f ) the MEK5/ERK5 inhibitor BIX02189, ( g ) ERK2 inhibitor TCS ERK 11e, and ( h ) ERK5 inhibitor ERK5-IN-1. Values are the mean ± SD ( n = 3). The hemolysis value corresponding to 0 μM of each antagonist and 4.6 μM rCPA was defined as 1. The statistical differences between each concentration of the inhibitor group and normal hemolysis group without inhibitor are indicated by letters a, b, c, and d, respectively, where “a” denotes p < 0.0001, “b” denotes p < 0.001, “c” denotes p < 0.01, “d” denotes p < 0.05, H denotes human red blood cells, and M denotes mouse red blood cells.

Article Snippet: Pyridoxal phosphate-6-azo (benzene-2,4-disulfonic acid) tetrasodium salt hydrate (PPADS), Suramin, MRS2179, Brilliant Blue G(BBG), ATP-2′, 3′-dialdehyde (OxATP), SL327, carbenoxolone disodium salt, mefloquine, apyrase, BIRB 796, DPI (Diphenyleneiodonium), and BIX02189 were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Concentration Assay