Journal: Antibiotics

Article Title: Kinase Inhibitor Library Screening Identifies the Cancer Therapeutic Sorafenib and Structurally Similar Compounds as Strong Inhibitors of the Fungal Pathogen Histoplasma capsulatum

doi: 10.3390/antibiotics10101223

Figure Lengend Snippet: Graphical representation of primary screen results of the Published Kinase Inhibitor Set ( A ) and the SelleckChem MAPK Inhibitor Library ( B ) against Histoplasma capsulatum . Each individual compound was tested, in duplicate, at a final concentration of 5 μM. Log phase Hc WU15 yeasts were seeded in 96 well plates at a density of 1 × 10 6 yeasts/mL in HMM/uracil. Compounds were diluted in DMSO and added to each well at a final DMSO concentration of 0.5%. Plates were incubated at 37 °C and 5% CO 2 with twice-daily shaking to improve aeration. OD 595 readings were taken on day 0 and day 4. The results for each compound are shown as average percent growth inhibition over the four day time period relative to the DMSO control. All compounds showing greater than 50% growth inhibition at day 4 are identified.

Article Snippet: The PKIS library and the MAP kinase inhibitor library L3400 (SelleckChem, Houston, TX, USA) were provided as 1 mM stocks in DMSO.

Techniques: Concentration Assay, Incubation, Inhibition, Control

Journal: World Journal of Gastroenterology : WJG

Article Title: Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line

doi: 10.3748/wjg.v10.i23.3522

Figure Lengend Snippet: Confirmation of the successful transfection of HBx into HepG2 cells by RT-PCR. Lane M: markers (the brightest band of the marker presents the 500 bp); lane A: transfected cells; lane B: transfected cells treated with U0126; lane C: the control.

Article Snippet: Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), lipofectamine 2 000 and G418 were purchased from Invitrogen Inc. ERK/MAPK pathway inhibitor U0126, was purchased from Alexis Inc. Anti-phospho-ERK 1 /ERK 2 rabbit polyclonal antibody was purchased from RD systems.

Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction, Marker

Journal: World Journal of Gastroenterology : WJG

Article Title: Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line

doi: 10.3748/wjg.v10.i23.3522

Figure Lengend Snippet: Apoptosis index of three different groups of cells after treatment with 5-Fu. A: HepG2/pcDNA3 cells; B: HepG2/ pcDNA3-HBx cells; C: U0126 pretreated transfected cells. D: Apoptotic index quantitated by FACS.

Article Snippet: Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), lipofectamine 2 000 and G418 were purchased from Invitrogen Inc. ERK/MAPK pathway inhibitor U0126, was purchased from Alexis Inc. Anti-phospho-ERK 1 /ERK 2 rabbit polyclonal antibody was purchased from RD systems.

Techniques: Transfection

Journal: World Journal of Gastroenterology : WJG

Article Title: Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line

doi: 10.3748/wjg.v10.i23.3522

Figure Lengend Snippet: Protein levels of MDR associated genes in different groups of cells. Lane1: HepG2/pcDNA3 cells; lane2: HepG2/pcDNA3-HBx cells; lane3: transfected cells treated with U0126. The primary antibody for A, B, C, D was β -actin, MDR-1, MRP-1 and LRP respectively.

Article Snippet: Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), lipofectamine 2 000 and G418 were purchased from Invitrogen Inc. ERK/MAPK pathway inhibitor U0126, was purchased from Alexis Inc. Anti-phospho-ERK 1 /ERK 2 rabbit polyclonal antibody was purchased from RD systems.

Techniques: Transfection

Journal: World Journal of Gastroenterology : WJG

Article Title: Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line

doi: 10.3748/wjg.v10.i23.3522

Figure Lengend Snippet: Increased phosphorylation of ERK induced by HBx in hepatoma cells analyzed by Western blot. Lane 1: trans-fected cells after treated with U0126; lane 2: HBx-expressing cells ; lane 3: control cells. A: Specific antibody for phosphory-lated-ERK; B: Specific antibody for total ERK.

Article Snippet: Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), lipofectamine 2 000 and G418 were purchased from Invitrogen Inc. ERK/MAPK pathway inhibitor U0126, was purchased from Alexis Inc. Anti-phospho-ERK 1 /ERK 2 rabbit polyclonal antibody was purchased from RD systems.

Techniques: Western Blot, Expressing

Journal: World Journal of Gastroenterology : WJG

Article Title: Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line

doi: 10.3748/wjg.v10.i23.3522

Figure Lengend Snippet: Changes of MDR related genes after treated with U0126 (mean ± SD)

Article Snippet: Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), lipofectamine 2 000 and G418 were purchased from Invitrogen Inc. ERK/MAPK pathway inhibitor U0126, was purchased from Alexis Inc. Anti-phospho-ERK 1 /ERK 2 rabbit polyclonal antibody was purchased from RD systems.

Techniques:

Journal: World Journal of Gastroenterology : WJG

Article Title: Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line

doi: 10.3748/wjg.v10.i23.3522

Figure Lengend Snippet: Changes of MDR related proteins after treated with U0126 (mean ± SD)

Article Snippet: Reagents Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), lipofectamine 2 000 and G418 were purchased from Invitrogen Inc. ERK/MAPK pathway inhibitor U0126, was purchased from Alexis Inc. Anti-phospho-ERK 1 /ERK 2 rabbit polyclonal antibody was purchased from RD systems.

Techniques:

Journal: Journal of Hematology & Oncology

Article Title: Triple MAPK inhibition salvaged a relapsed post-BCMA CAR-T cell therapy multiple myeloma patient with a BRAF V600E subclonal mutation

doi: 10.1186/s13045-022-01330-3

Figure Lengend Snippet: Triple MAPK inhibition effectively reduces phosphorylated ERK in BRAF (V600E) CD138 + plasma cells A WB of magnetic bead selected CD138 + plasma cells from RRMM patient’s BMA after 48 h in vitro treatment with encorafenib (ENC; 50 nM) and binimetinib (BIN; 250 nM), regorafenib alone (REG; 1μΜ), or combination of the three drugs. B Travera analysis on RRMM patient CD138 + cells showing sensitivity to trametinib (TRAM) in combination with dabrafenib (DAB) and regorafinib (REG) at varying concentrations. C Relative pERK protein expression after quantification and normalization to actin

Article Snippet: RRMM patient’s BMA selected CD138 + cells’ sensitivity to MAPK inhibitors were assessed by Travera.

Techniques: Inhibition, Clinical Proteomics, In Vitro, Expressing

Journal: Journal of Hematology & Oncology

Article Title: Triple MAPK inhibition salvaged a relapsed post-BCMA CAR-T cell therapy multiple myeloma patient with a BRAF V600E subclonal mutation

doi: 10.1186/s13045-022-01330-3

Figure Lengend Snippet: Temporal evolution and trajectory of MAPK alterations. A Clusters of subclonal mutations sampled over time with the mutational VAF represented as a percentage on the y axis show the trajectory of the subclone harboring the BRAF V600E mutation over time in response to treatment and the later emergence of a subclonal KRAS Q61R mutation after the end of triple MAPK inhibition. B A reconstructed phylogenetic tree of subclones across all time points shows that the BRAF V600E is subclonal and ancestral to the clone that gives rise to a KRAS Q61R mutation. C RNA expression shows an increase in MAPK pathway activation, measured as a combined z-score, over time in response to treatment with triple MAPK inhibition therapy

Article Snippet: RRMM patient’s BMA selected CD138 + cells’ sensitivity to MAPK inhibitors were assessed by Travera.

Techniques: Mutagenesis, Inhibition, RNA Expression, Activation Assay

Journal: Journal of Hematology & Oncology

Article Title: Triple MAPK inhibition salvaged a relapsed post-BCMA CAR-T cell therapy multiple myeloma patient with a BRAF V600E subclonal mutation

doi: 10.1186/s13045-022-01330-3

Figure Lengend Snippet: CD138 + BMA plasma cells shift their dependency to PI3K/AKT pathway with increased sensitivity to copanlisib A WB of magnetic bead selected CD138 + plasma cells from RRMM patient’s BMA after 48 h in vitro treatment with with encorafenib (ENC; 50 nM) and binimetinib (BIN; 250 nM), regorafenib alone (REG; 1μΜ), combination of the three drugs or copanlisib alone (25 nM). B Rlative protein expression of pERK, pS6, pAKT and BRAF (V600E) after quantification and normalization to actin C CD138 + viability measurement after 48 h in vitro treatment with with encorafenib (ENC; 50 nM) and binimetinib (BIN; 250 nM), regorafenib alone (REG; 1 μΜ), combination of the three drugs or copanlisib alone (25 nM). D RNA expression shows elevated PI3K/Akt pathway activation, measured as a combined z-score, throughout the course of treatment with triple MAPK inhibition therapy

Article Snippet: RRMM patient’s BMA selected CD138 + cells’ sensitivity to MAPK inhibitors were assessed by Travera.

Techniques: Clinical Proteomics, In Vitro, Expressing, RNA Expression, Activation Assay, Inhibition