mapk inhibitors pd98059  (Millipore)

Journal: bioRxiv

Article Title: Targeting one-carbon metabolic vulnerabilities of metastasis with therapeutic potential

doi: 10.64898/2026.02.07.704548

Figure Lengend Snippet: ( A ) Schematic of the metastasis functional screening using molecular inhibitors in vivo. ( B, C, and D ) Gross pulmonary metastases from human melanoma A375sm (B), mouse melanoma B16F10 (C), and mouse rhabdomyosarcoma RMS14 (D) in response to molecular inhibitor in the experimental metastasis assay by tail vein injection. Data are represented as mean ± SEM. The p-values were presented from an unpaired t-test analysis (two-tailed) compared with the control (DMSO). Cal C, 100 nM calphostin C; Rho, 12nM Rho Inhibitor I; Rap, 10nM Rapamycin; H-89, 50nM PKA inhibitor; SB, 10µM p38 inhibitor SB203580; TSA, 300nM HDAC inhibitor Trichostatin A; MS, 10µM HDAC inhibitor MS-275; PD, 20µM MAPK inhibitor PD98059; LY, 10µM PI3K/AKT inhibitor LY294002. ND , no statistical difference; * p < 0.05; ** p < 0.01; *** p < 0.001. n = 10. ( E, F, and G ) The related cell viability of human melanoma A375sm (E), mouse melanoma B16F10 (F), and mouse rhabdomyosarcoma (G) cells, pretreated with molecular inhibitors. Data are represented as mean ± SEM of three independent experiments. The p-values were presented from an unpaired t-test analysis (two-tailed) compared with the control (DMSO). ND , no statistical difference; * p < 0.05; ** p < 0.01; *** p < 0.001. ( H ) Gross pulmonary metastases from mouse melanoma B16F10 pretreated with 10µM PI3K/AKT inhibitor LY294002 at the indicated time (LY12, 12 hours, LY24, 24 hours and LY48, 48 hours). Data are represented as mean ± SEM. The p-values were presented from an unpaired t-test analysis (two-tailed) compared with the control (DMSO). ND , no statistical difference; * p < 0.05; ** p < 0.01; *** p < 0.001. n=10. ( I ) Gross pulmonary metastases from mouse rhabdomyosarcoma RMS14 cells pretreated with 20µM MAPK inhibitor PD98059 at the indicated time (PD12, 12 hours, PD24, 24 hours and PD48, 48 hours). Data are represented as mean ± SEM. The p-values were presented from an unpaired t-test analysis (two-tailed) compared with the control (DMSO). ND , no statistical difference; * p < 0.05; ** p < 0.01; *** p < 0.001. n=10.

Article Snippet: Calphostin C, Rho Inhibitor I, Rapamycin, PKA inhibitor H-89, p38 inhibitor SB203580, HADC inhibitor Trichostatin A and MS-275, MAPK inhibitor PD98059, PI3K/AKT inhibitor LY294002, Rottlerin, bisindoylmaleimide I (BMI), PKC β inhibitor I, and FDA-approved-Drug-Library (1562 compounds, HY-L022) were obtained from MedChemExpress (Monmouth Junction, NJ).

Techniques: Functional Assay, In Vivo, Injection, Two Tailed Test, Control

Journal: bioRxiv

Article Title: Targeting one-carbon metabolic vulnerabilities of metastasis with therapeutic potential

doi: 10.64898/2026.02.07.704548

Figure Lengend Snippet: (A) Schematic of microarray analysis to focus on the inhibition of tumor metastasis. (B) The significant differentiated gene expression patterns associated with the inhibition of tumor metastasis progression and the identification of metabolism pathways associated with tumor metastasis. The significantly expressed genes in B16F10 (a) or RMS14 (b) cells treated with inhibitors (Cal C, 100 nM calphostin; Rap, 10nM Rapamycin; 20µM MAPK inhibitor PD98059; LY, 10µM PI3K/AKT inhibitor LY294002) compared with DMSO control were filtered by a p-value of 0.05 and an absolute value of fold change of 1.5 in ANOVA analysis. a1, Cal C v.s. DMSO; a2, Rap v.s. DMSO; a3, LY24 v.s. DMSO; a4, LY48 v.s. DMSO in B16F10 cells. b1, Cal C v.s. DMSO; b2, Rap v.s. DMSO; b3, PD24 v.s. DMSO; b4, PD48 v.s. DMSO in RMS14 cells. ( C ) Gene signaling pathway (GO gene pathway) analysis revealed that top 15 biological pathways and functions within a given gene list from micrroarry analysis regulate one-carbon metabolism and de novo serine synthesis (SSP). Both p and FDR valuest were transformed to negative log (-log2) values. ( D ) Schematic of the one-carbon metabolism and De novo serine synthesis (SSP) pathways. ( E to H ) Validation of gene expression identified in cDNA microarray analysis by Western blot analysis. The human melanoma A375sm (E) and WM88 (F), mouse melanoma B16F10 (G), or mouse rhabdomyosarcoma RMS14 (H) cells were treated with molecular inhibitors. DMSO, as a mock control; Cal C, 100 nM calphostin C; Rho, 12nM Rho Inhibitor I; Rap, 10nM Rapamycin; SB, 10 µM p38 inhibitor SB203580; PD, 20 µM MAPK inhibitor PD98059; LY, 10 µM PI3K/AKT inhibitor LY294002; Rott, 5μM of rottlerin; BMI, 20 nM of bisindoylmaleimide I; Iβ, 20 nM of PKCβ inhibitor I. β -actin was used as a control.

Article Snippet: Calphostin C, Rho Inhibitor I, Rapamycin, PKA inhibitor H-89, p38 inhibitor SB203580, HADC inhibitor Trichostatin A and MS-275, MAPK inhibitor PD98059, PI3K/AKT inhibitor LY294002, Rottlerin, bisindoylmaleimide I (BMI), PKC β inhibitor I, and FDA-approved-Drug-Library (1562 compounds, HY-L022) were obtained from MedChemExpress (Monmouth Junction, NJ).

Techniques: Microarray, Inhibition, Gene Expression, Control, Transformation Assay, Biomarker Discovery, Western Blot

Journal: bioRxiv

Article Title: Targeting one-carbon metabolic vulnerabilities of metastasis with therapeutic potential

doi: 10.64898/2026.02.07.704548

Figure Lengend Snippet: ( A ) Western blotting showed the expression of the indicated one-carbon and SSP metabolism pathway genes treated with different compounds, including identified new compounds comp4, comp7, and comp9. The cells were treated with 5 µM NCT503, 10 µM comp2, 10 µM comp3, 100 nM comp4, 10µM venetoclax, 10 µM comp6, 10 nM comp7, 30 nM CB-839, 10µM comp9, DMSO, 5 mM 2-DG, 1 mM BSO, 10 µM comp13, 10 µM comp14, 2 mM metf (Metformin) for 36 hours. ( B, C ) Seahorse analysis showed the effect of the different compounds on OCR (B) and ATP production (C). The cells were treated with 100 nM calphostin (Cal C), 10nM Rapamycin (Rap), 20µM MAPK inhibitor PD98059 (PD), 10µM PI3K/AKT inhibitor LY294002 (LY), 100 nM comp4, 5 µM NCT503, 10 nM comp7, 10µM comp9, 2 mM Metformin(Metf), 10 µM comp13, 10 µM comp14 and DMSO control. ND , no statistical difference; * p < 0.05; ** p < 0.01; *** p < 0.001. ( D, E ) Representation of bioluminescence signals of tumor metastasis from before treatment (D) and treated for two weeks with indicated compouds (E) after A375sm cell transplantation in NSG mice tracked by of the In Vivo Imaging System (IVIS). C, DMSO control; NCT503, 20 mg/Kg; Comp2, 25 mg/Kg; Comp4, 30 mg/Kg; comp7, 3 mg/Kg; comp9, 10 mg/Kg; venetoclax, 12 mg/Kg. ( F ) Gross pulmonary metastases from mice transplanted with cells by tail vein injection and treated with different compounds. N=10. Graphs show the mean ± SEM. The p-value is shown by an unpaired t-test (two-tailed). ND , no statistical difference; * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Calphostin C, Rho Inhibitor I, Rapamycin, PKA inhibitor H-89, p38 inhibitor SB203580, HADC inhibitor Trichostatin A and MS-275, MAPK inhibitor PD98059, PI3K/AKT inhibitor LY294002, Rottlerin, bisindoylmaleimide I (BMI), PKC β inhibitor I, and FDA-approved-Drug-Library (1562 compounds, HY-L022) were obtained from MedChemExpress (Monmouth Junction, NJ).

Techniques: Western Blot, Expressing, Control, Transplantation Assay, In Vivo Imaging, Injection, Two Tailed Test

Journal: Acta Pharmaceutica Sinica. B

Article Title: USP10-mediated deubiquitination and activation of KRAS mutants promotes colorectal cancer via a novel USP10/KRAS positive feedback circuit

doi: 10.1016/j.apsb.2025.11.015

Figure Lengend Snippet: KRAS mutants upregulates USP10 levels and stability by promoting the phosphorylation of USP10 at Tyr42 and Ser337. (A) KRAS -mutant CRC patients displayed higher USP10 levels compared to KRAS wild-type patients. Immunohistochemistry (IHC) staining of intratumor USP10 in KRAS mutants/wild-type CRC patients (mean ± SD, n = 6). (B) Overexpression of KRAS mutants significantly upregulated USP10 protein levels. HT-29 cells were transfected with the indicated plasmids and cell lysates were subjected to IB assays. (C) Overexpression of KRAS mutants, not KRAS wild-type, significantly promoted USP10 phosphorylation. HEK-293T cells transfected with indicated plasmids were immunoprecipitated with anti-Flag agarose beads and the protein remaining on anti-Flag agarose beads were subjected to IB assays. (D) MAPK inhibitor PD98059 reversed the upregulation of USP10 phosphorylation induced by KRAS mutants. HEK-293T cells transfected with indicated plasmids were treated with DMSO, PD98059 (10 μmol/L) for 24 h before harvested, then cell lysates were immunoprecipitated with anti-Flag agarose beads and immunoblotted with the indicated antibody. (E) T42 and S337 are the main phosphorylation sites of USP10 regulated by KRAS mutants. HEK-293T cells transfected with indicated plasmids were immunoprecipitated with anti-Flag agarose beads and the protein remaining on anti-Flag agarose beads was subjected to IB assays. TA&SA: T42A&S337A. (F) KRAS mutants significantly increased the protein levels and stability of USP10-WT, but not USP10-T42A&S337A. HT-29 cells expressing USP10-WT/TA&SA were infected with lentivirus encoding Vector or KRAS G12V -Myc and treated with 20 μg/mL CHX for indicated time and the cell lysates were immunoblotted with the indicated antibody. TA&SA: T42A&S337A. ∗∗ P < 0.01.

Article Snippet: Proteasome inhibitor MG132 (#S2619) and MAPK inhibitors PD98059 (#S1177) and SB203580 (#S1076) were obtained from Selleck (Houston, TX, USA).

Techniques: Phospho-proteomics, Mutagenesis, Immunohistochemistry, Over Expression, Transfection, Immunoprecipitation, Expressing, Infection, Plasmid Preparation

Journal: Antioxidants

Article Title: NADPH Oxidase 5 (NOX5) Upregulates MMP-10 Production and Cell Migration in Human Endothelial Cells

doi: 10.3390/antiox13101199

Figure Lengend Snippet: p38 MAPK, MEK/ERK, and JNK pathways participate at different levels in NOX5-dependent regulation of MMP-10: ( A ) MMP-10 mRNA levels of tHMock and tHNOX5 cell lines after 24 h of incubation with 5 μM 219138-24-6 (p38 MAPK inhibitor, ip38), PD98059 (MEK/ERK inhibitor, iERK), or JNK-IN-8 (JNK inhibitor, iJNK) ( n = 6). ( B ) MMP-10 protein in the conditioned medium of tHMock and tHNOX5 cell lines after 24 h of incubation with 5 μM 219138-24-6 (ip38), PD98059 (iERK), or JNK-IN-8 (iJNK) ( n = 6). tHMock: stable cell line transfected with pcDNA3.2-Mock. tHNOX5: stable cell line transfected with pcDNA3.2-NOX5. n.s.: not significant differences, * p < 0.05, *** p < 0.001, **** p < 0.0001. Data are presented as median and IQR.

Article Snippet: The p38 MAPK inhibitor (ab145872, Abcam ® , Waltham, MA, USA), JNK MAPK inhibitor (JNK-IN-8, SML1246, Sigma Aldrich), and ERK MAPK inhibitor (PD98059, 9900S, Cell Signaling Technology, Danvers, MA, USA) were used at final concentrations of 5 μM in the cell medium.

Techniques: Incubation, Stable Transfection, Transfection

Journal: Antioxidants

Article Title: NADPH Oxidase 5 (NOX5) Upregulates MMP-10 Production and Cell Migration in Human Endothelial Cells

doi: 10.3390/antiox13101199

Figure Lengend Snippet: NOX5 enhances MMP-10 promoter activity via the JNK pathway and a functional AP-1 site: ( A ) MMP-10 promoter activity of tHMock and tHNOX5 cell lines at baseline and stimulated with 0.25 μM Ang II ( n = 6). ( B ) MMP-10 promoter activity of tHMock and tHNOX5 cell lines after 24 h of incubation with 5 μM 219138-24-6 (p38 MAPK inhibitor, ip38), PD98059 (MEK/ERK inhibitor, iERK), or JNK-IN-8 (JNK inhibitor, iJNK) ( n = 6). ( C ) Schematic representation of MMP-10 promoter constructions used and quantification of MMP-10 promoter activity of tHMock and tHNOX5 cell lines transfected with MMP-10 promoter constructions ( n = 6). CREB: cAMP response element binding protein (CREB) putative union site. AP-1: activator protein-1 (AP-1) putative union site. Crosses indicate site-directed mutations in the putative union sites. tHMock: stable cell line transfected with pcDNA3.2-Mock. tHNOX5: stable cell line transfected with pcDNA3.2-NOX5. ns: not significant differences, * p < 0.05 vs. tHMock Ctrl, *** p < 0.001, **** p < 0.0001, # p < 0.05 vs. tHNOX5 Ctrl cells. Data are presented as median and IQR or as mean ± SEM.

Article Snippet: The p38 MAPK inhibitor (ab145872, Abcam ® , Waltham, MA, USA), JNK MAPK inhibitor (JNK-IN-8, SML1246, Sigma Aldrich), and ERK MAPK inhibitor (PD98059, 9900S, Cell Signaling Technology, Danvers, MA, USA) were used at final concentrations of 5 μM in the cell medium.

Techniques: Activity Assay, Functional Assay, Incubation, Transfection, Binding Assay, Stable Transfection