human gas6 hgas6  (R&D Systems)

Journal: NPJ Precision Oncology

Article Title: AXL–SHC1 signaling axis mediates adaptive resistance to HER2-targeted tyrosine kinase inhibitors in HER2-aberrant lung and gastric cancers

doi: 10.1038/s41698-026-01385-2

Figure Lengend Snippet: A qPCR analysis of GAS6 expression in Calu-3 and MKN7 parent cells treated with nonspecific (control) or GAS6-specific siRNAs for 48 h. * P < 0.05 (unpaired t-tests). B MTT assays evaluating cell viability of Calu-3 and MKN7 cells treated with nonspecific or GAS6-specific siRNAs were incubated with or without mobocertinib (0.01 or 0.1 μmol/L) for 72 h. These cell lines were incubated in an RPMI 1640 medium with 0.1% fetal bovine serum, penicillin (100 U/mL), and streptomycin (100 μg/mL) in a humidified 5% CO2 incubator at 37 °C for 72 h (* P < 0.05, one-way ANOVA). C Western blotting analysis of Calu-3 and MKN7 cells transfected with nonspecific or GAS6-specific siRNAs for 48 h, followed by treatment with or without mobocertinib (0.01 or 0.1 μmol/L) for 4 h. D Calu-3 and MKN7 cells with nonspecific siRNA or AXL-specific siRNAs were treated for 72 h with or without mobocertinib (0.01 or 0.1 μmol/L) and/or GAS6 (50 ng/mL). Cell growth was determined using MTT assays. * P < 0.05 (two-way ANOVA). E Western blotting of Calu-3 and MKN7 cells transfected with nonspecific or AXL-specific siRNAs and treated with or without mobocertinib (0.01 or 0.1 μmol/L) for 4 h and GAS6 (50 ng/mL) for 15 min. F MKN7 cells treated with or without mobocertinib (Mobo, 0.1 μmol/L) for 24 h were detected by western blotting with immunoprecipitation of the indicated proteins.

Article Snippet: Recombinant human GAS6 was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Control, Incubation, Western Blot, Transfection, Immunoprecipitation

Journal: NPJ Precision Oncology

Article Title: AXL–SHC1 signaling axis mediates adaptive resistance to HER2-targeted tyrosine kinase inhibitors in HER2-aberrant lung and gastric cancers

doi: 10.1038/s41698-026-01385-2

Figure Lengend Snippet: A Combination of mobocertinib and knockdown of several adapter proteins in Calu-3. and MKN7 cells. Calu-3 and MKN7 cells treated with nonspecific control, FYN, GRB2, PTPN11, Shc1, or SRC-specific siRNAs were incubated with or without mobocertinib (0.01 or 0.1 μmol/L) for 72 h, and cell viability was detected using MTT assays. * P < 0.05 compared with nonspecific control siRNA (two-way ANOVA). Data are represented as the mean ± S.D. B Western blotting of Calu-3 and MKN7 cells treated with nonspecific control siRNA or Shc1-specific siRNA and incubated with or without mobocertinib (0.01 or 0.1 μmol/L) for 24 h. C Calu-3 and MKN7 cells treated with or without mobocertinib (Mobo; 0.01 or 0.1 μmol/L) for 24 h were detected by western blotting with immunoprecipitation of the indicated proteins. D Western blotting showing nuclear localization of ShcBP1 in Calu-3 and MKN7 cells treated with mobocertinib (0.01 or 0.1 μmol/L) for 72 h. E Immunofluorescence showing nuclear localization of ShcBP1 in Calu-3 and MKN7 cells treated with mobocertinib (0.01 or 0.1 μmol/L) for 72 h. F MTT assays assessing cell viability of Calu-3 and MKN7 treated with nonspecific siRNA or ShcBP1-specific siRNAs, and incubated with or without mobocertinib * P < 0.05 (one-way ANOVA). G Schematic diagram illustrating the drug tolerance mechanisms, including GAS6–AXL activation through the SHC1BP–SHC1 complex, in HER2-aberrant cancer cells.

Article Snippet: Recombinant human GAS6 was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Knockdown, Control, Incubation, Western Blot, Immunoprecipitation, Immunofluorescence, Activation Assay