mitogen activated protein kinase mapk inhibitors  (MedChemExpress)

Journal: Journal of Cellular and Molecular Medicine

Article Title: The Efficacy of Hellebrigenin Against Nasopharyngeal Carcinoma Cells: The Molecular and Bioinformatic Analysis

doi: 10.1111/jcmm.70624

Figure Lengend Snippet: Effects of hellebrigenin on apoptosis in NPC cells and in an NPC xenograft model. (A, B) MAPK protein expression was assessed using a Western blot assay following hellebrigenin treatment (0, 5, 10 and 20 nM). β‐Actin was identified as the internal control. The results are presented as mean ± SD, n = 3. * p < 0.05 compared with 0 nM. (C, D) Cells were treated with 10 nM hellebrigenin with or without the ERK inhibitor U0126. β‐Actin was identified as the internal control. The results are presented as mean ± SD, n = 3. * p < 0.05 compared with 0 nM. # p < 0.05 compared with hellebrigenin 10 nM. (E, F) Cells were treated with 10 nM of hellebrigenin with or without the JNK inhibitor JNK‐in‐8. β‐Actin was identified as the internal control. The results are presented as mean ± SD, n = 3. * p < 0.05 compared with 0 nM. # p < 0.05 compared with hellebrigenin 10 nM. (G) Tumour volumes of the vehicle group and NPC xenograft mice treated with 4 mg/kg of hellebrigenin. The seventh day following tumour injection was identified as Day 0. The results are presented as mean ± SE, n = 3. * p < 0.05 compared with vehicle. (H) Body weights of the vehicle group and NPC xenograft mice treated with 4 mg/kg of hellebrigenin. The results are presented as mean ± SE, n = 3. * p < 0.05 compared with vehicle. (I) Immunohistochemical analysis of Ki67 and CHCHD2 expression in the vehicle group and NPC xenograft mice treated with 4 mg/kg of hellebrigenin. Scale bar = 50 μm.

Article Snippet: Hellebrigenin (purity ≥ 99%, Figure ) and mitogen‐activated protein kinase (MAPK) inhibitors (U0126 and JNK‐in‐8) were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Expressing, Western Blot, Control, Injection, Immunohistochemical staining

Journal: Biomolecules & Therapeutics

Article Title: Cynaropicrin Induces Reactive Oxygen Species-Dependent Paraptosis-Like Cell Death in Human Liver Cancer Cells

doi: 10.4062/biomolther.2025.011

Figure Lengend Snippet: Alleviation of cynaropicrin-induced cytotoxicity through a p38 MAPK inhibitor in Hep3B cells. (A) After treating cells with cynaropicrin for the indicated duration, changes in the expression of the three types of MAPKs were examined via Western blot analysis. (B-D) Cells were pretreated with the p38 MAPK inhibitor SB203580 for 1 h and then treated with cynaropicrin for 24 h. Thereafter, changes in morphology (B), cell viability (C), and expression of the indicated proteins (D) were examined. (C) Numerical data are presented as mean ± SD (n=3). *** p <0.001 compared to untreated group; ### p <0.001 compared to cynaropicrin-treated group.

Article Snippet: Antibodies against poly(ADP-ribose) polymerase (PARP), apoptosis-linked gene 2-interacting protein X (Alix), p38 mitogen activated protein kinase (MAPK), extracellular signal-regulated kinase ( de Ridder et al ., 2023 ), c-Jun N-terminal kinases (JNK), phosphor (p)-JNK, and β-actin were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).

Techniques: Expressing, Western Blot

Journal: Biomolecules & Therapeutics

Article Title: Cynaropicrin Induces Reactive Oxygen Species-Dependent Paraptosis-Like Cell Death in Human Liver Cancer Cells

doi: 10.4062/biomolther.2025.011

Figure Lengend Snippet: Effects of the p38 MAPK inhibitor on cynaropicrin-induced ER stress and mitochondrial impairment in Hep3B cells. The effects of SB203580 on the cynaropicrin-induced increase in intracellular Ca 2+ concentration (A, B), ER stress (C, D), and mitochondrial dysfunction (E, F) were evaluated via flow cytometry. (B, D, F) Numerical data are presented as mean ± SD (n=3). *** p <0.001 compared to untreated group; # p <0.05 and ### p <0.001 compared to cynaropicrin-treated group.

Article Snippet: Antibodies against poly(ADP-ribose) polymerase (PARP), apoptosis-linked gene 2-interacting protein X (Alix), p38 mitogen activated protein kinase (MAPK), extracellular signal-regulated kinase ( de Ridder et al ., 2023 ), c-Jun N-terminal kinases (JNK), phosphor (p)-JNK, and β-actin were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).

Techniques: Concentration Assay, Flow Cytometry

Journal: Biomolecules & Therapeutics

Article Title: Cynaropicrin Induces Reactive Oxygen Species-Dependent Paraptosis-Like Cell Death in Human Liver Cancer Cells

doi: 10.4062/biomolther.2025.011

Figure Lengend Snippet: A schematic diagram illustrating the mechanisms by which cynaropicrin induces cell death in Hep3B cells, including the promotion of mitochondrial ROS production, activation of the p38 MAPK pathway, inhibition of Alix protein, and enhancement of ER stress.

Article Snippet: Antibodies against poly(ADP-ribose) polymerase (PARP), apoptosis-linked gene 2-interacting protein X (Alix), p38 mitogen activated protein kinase (MAPK), extracellular signal-regulated kinase ( de Ridder et al ., 2023 ), c-Jun N-terminal kinases (JNK), phosphor (p)-JNK, and β-actin were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).

Techniques: Activation Assay, Inhibition

Journal: American journal of hematology

Article Title: RAS signaling pathway is essential in regulating PIEZO1-mediated hepatic iron overload in dehydrated hereditary stomatocytosis.

doi: 10.1002/ajh.27523

Figure Lengend Snippet: FIGURE 4 RRAS is a key regulator of PIEZO1-mediated HAMP gene regulation, modulating MAPK cascade and RAS-mediated signaling. (A) Representative immunoblots of pERK1/2 in total cells lysate of PIEZO1-WT and PIEZO1-KI cells treated with Yoda-1 (15 μM for 60 min), PIEZO1-KI treated with Yoda-1 plus GsMTx-4 (5 μM for 30 min) and PIEZO1-KI treated with U0126 for 30 and 60 min. tERK1/2 was used as the loading control. (B) Histograms showing HAMP gene expression in PIEZO1-WT and PIEZO1-KI treated with Yoda-1 (15 μM for 60 min), PIEZO1-KI treated with Yoda-1 plus GsMTx-4 (5 μM for 30 min), and PIEZO1-KI treated with U0126 for 30 and 60 min. Scatter dot plot showing quantification by densitometric analysis of three separate western blots with similar results. Data are means ± standard deviation (SD) of three independent experiments (*p < .05, **p < .01, ANOVA test and post hoc correction by Sidak's multiple comparison tests). (C) Left panel: Representative immunoblots of RAS GTP proteins purified by affinity precipitation with the RAS binding domain (RBD) of RAF1 from total cell lysate of PIEZO1-WT and PIEZO1-KI cells at steady state and PIEZO1-KI treated with Yoda-1 (15 μM for 60 min) or Yoda-1 plus GsMTx-4 (5 μM for 30 min). Lanes 5 and 6 are the positive controls (cell lysate treated with GST-Raf1-RBD plus GDP or GTP); lane 7 is the negative control (without GST-Raf1-RBD). Right panel: Quantification by densitometric analysis of three separate western blots with similar results. Data are means ± SD (*p < .05, PIEZO1-WT vs. PIEZO1-KI; **p < .01, PIEZO1-KI vs. PIEZO1-KI + GsMTx-4; ANOVA test and post hoc correction by Sidak's multiple comparison tests). (D) Left panel: Representative immunoblots of RAS GTP proteins purified by affinity precipitation with the RBD of RAF1 from total cell lysate of primary murine hepatocytes (HCs) treated with vehicle, Yoda-1 (50 μM for 30 min) or Yoda-1 plus GsMTx-4 (30 μM for 30 min). Lanes 5 and 6 are the positive controls (cell lysate treated with GST-Raf1-RBD plus GDP or GTP); lane 7 is the negative control (without GST-Raf1-RBD). Right panel: Histograms showing quantification by densitometric analysis of three separate western blots with similar results. Data are means ± SD (*p < .05, hepatocytes + Yoda-1 vs. hepatocytes vehicle-treated by ANOVA test and post hoc correction by Sidak's multiple comparison tests). (E) Left panel: Representative immunoblots of RAS GTP proteins purified by affinity precipitation with the RBD of RAF1 from total cell lysate of PIEZO1-WT and PIEZO1-KI cells at steady state and PIEZO1-KI treated with Yoda-1, Yoda-1 plus GsMTx-4 or Salirasib 25 and 100 μM. Right panel: Histograms showing HAMP gene expression in PIEZO1-WT and PIEZO1-KI treated with Yoda-1 (15 μM for 60 min), PIEZO1-KI treated with Yoda-1 plus GsMTx-4 and PIEZO1-KI treated with Salirasib 25 and 100 μM. Boxes showing quantification by densitometric analysis of three separate Western blots with similar results. Data are means ± SD of three independent experiments. (*p < .05, PIEZO1-WT vs. PIEZO1-KI; **p < .01, PIEZO1-KI vs. PIEZO1-KI + GsMTx-4; ANOVA test and post hoc correction by Sidak's multiple comparison tests). (G) Left panel: Representative immunoblots of RRAS, and ferritin in total cells lysate of PIEZO1-WT and PIEZO1-KI cells treated with Holo-transferrin (Holo-Tf) (30 μM) for 0, 2, 4, or 16 h. GAPDH is the loading control. Right panel: Quantification by densitometric analysis of three separate Western blots with similar results. Data are means ± SD (*p < .05, **p < .01 PIEZO1-WT vs. PIEZO1-KI; ##p < .01 PIEZO1-KI 2, 4, or 6 vs. PIEZO1-KI NT [0 h]; ##p < .01 PIEZO1-KI vs. PIEZO1-WT. ANOVA test and post hoc correction by Sidak's multiple comparison tests). (H) Histograms showing HAMP gene expression normalized to β-actin (red), and scatter dot plot showing quantification of and RRAS protein expression in PIEZO1-KI cells treated with Holo-Tf (30 μM) for 2, 4, or 16 h. Data are means ± SD of three independent experiments and are represented as fold on NT (##p < .01 vs. NT (0 h);°°p < .01, PIEZO1-KI 2 h vs. PIEZO1-KI 4 h, PIEZO1-KI 4 h vs. PIEZO1-KI 16 h ANOVA and PIEZO1-KI 2 h vs. PIEZO1-KI 16 h by ANOVA test and post hoc correction by Sidak's for multiple comparisons). OD, optical density.

Article Snippet: U0126 dual specificity mitogen-activated protein kinase kinase 1 and 2 (MEK1/2) inhibitor; Cat. N° 1144; Bio-techne TOCRIS) was added to PIEZO1-WT and PIEZO1-KI cells at 10 μM for 30 and 60 min.

Techniques: Western Blot, Control, Gene Expression, Standard Deviation, Comparison, Purification, Affinity Precipitation, Binding Assay, Negative Control, Expressing

Journal: BMC cancer

Article Title: Aldo-keto reductase family 1 member C3 mediates radioresistance of esophageal cancer cells through suppressing MAPK and AKT signaling.

doi: 10.1186/s12885-024-13012-z

Figure Lengend Snippet: Fig. 4 The effect of AKR1C3, MEK, or AKT inhibition on colony formation of KYSE170R cells. KYSE170R cells were treated with the inhibitor of AKR1C3 (AKR1C3-IN-1, 20 μM), MEK (PD98059, 20 μM), AKT (MK-2206, 20 μM), or JNK (SP600125, 20 μM) for 24 h, followed by exposure to different doses of 6 MV X-rays (0, 2, 4, and 8 Gy). The irradiated cells were collected, seeded in 6-well plates at 500 (0 Gy group), 1000 (2 and 4 Gy groups), or 2000 (8 Gy group) cells/well, and cultured for 7–10 days. The numbers of colonies were counted. The surviving fraction was calculated. Representative images are shown. Magnification 100 ×. Data are expressed as the mean ± SD

Article Snippet: To investigate the roles of MAPK and AKT signaling in AKR1C3-regulated radiosensitivity of KYSE170R cells, we treated KYSE170R cells with AKR1C3 inhibitor AKR1C3-IN-1 (20 μM; MedChemExpress, Monmouth Junction, NJ, USA), mitogen-activated protein kinase kinase (MEK) inhibitor PD98059 (20 μM; MedChemExpress), AKT inhibitor MK-2206 (20 μM; MedChemExpress), or JNK inhibitor SP600125 (20 μM; MedChemExpress) for 24 h, followed by exposure to different doses of 6 MV X-rays.

Techniques: Inhibition, Irradiation, Cell Culture

Journal: BMC cancer

Article Title: Aldo-keto reductase family 1 member C3 mediates radioresistance of esophageal cancer cells through suppressing MAPK and AKT signaling.

doi: 10.1186/s12885-024-13012-z

Figure Lengend Snippet: Fig. 5 The effect of MEK or AKT inhibition on colony formation of KYSE170R-shAKR1C3 cells. KYSE170R-shAKR1C3 cells were treated with the inhibitor of MEK (PD98059, 20 μM), AKT (MK-2206, 20 μM), or JNK (SP600125, 20 μM) for 24 h, followed by exposure to different doses of 6 MV X-rays (0, 2, 4, and 8 Gy). The irradiated cells were collected, seeded in 6-well plates at 500 (0 Gy group), 1000 (2 and 4 Gy groups), or 2000 (8 Gy group) cells/well, and cultured for 7–10 days. The numbers of colonies were counted. The surviving fraction was calculated. Representative images are shown. Magnification 100×. Data are expressed as the mean ± SD

Article Snippet: To investigate the roles of MAPK and AKT signaling in AKR1C3-regulated radiosensitivity of KYSE170R cells, we treated KYSE170R cells with AKR1C3 inhibitor AKR1C3-IN-1 (20 μM; MedChemExpress, Monmouth Junction, NJ, USA), mitogen-activated protein kinase kinase (MEK) inhibitor PD98059 (20 μM; MedChemExpress), AKT inhibitor MK-2206 (20 μM; MedChemExpress), or JNK inhibitor SP600125 (20 μM; MedChemExpress) for 24 h, followed by exposure to different doses of 6 MV X-rays.

Techniques: Inhibition, Irradiation, Cell Culture