Journal: Frontiers in Pharmacology

Article Title: Dasatinib suppresses particulate-induced pyroptosis and acute lung inflammation

doi: 10.3389/fphar.2023.1250383

Figure Lengend Snippet: Phosphorylated Src family kinases (p-SFKs) accumulate around silica particle (SP)-containing early phagosomes. Primed bone marrow-derived macrophages (BMDMs) were treated with dasatinib (20 µM) and stimulated or not stimulated with fluorescent SPs (1,500 nm in diameter, 300 μg/mL; cyan). Thirty minutes after stimulation with fluorescent SPs, p-SFKs (green) and RAB5A (magenta) were stained with specific antibodies. Yellow arrowheads indicate SPs surrounded by both p-SFKs and RAB5A. Scale bar; 10 µm.

Article Snippet: Anti-phospho-Src family (D49G4), anti-RAB5A, member RAS oncogene family (RAB5A) (E6N8S), HRP-conjugated anti-mouse IgG, and HRP-conjugated anti-rabbit IgG antibodies were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Derivative Assay, Staining

Journal: International Journal of Molecular Sciences

Article Title: AMPK/FOXO3a Pathway Increases Activity and/or Expression of ATM, DNA-PKcs, Src, EGFR, PDK1, and SOD2 and Induces Radioresistance under Nutrient Starvation

doi: 10.3390/ijms241612828

Figure Lengend Snippet: Effects of nutrient starvation on clonogenic cell survival and phosphorylation and/or expression of AMPKα, ATM, DNA-PKcs, FOXO3a, Src, EGFR, PDK1, SOD2, and HIF-1α in MDA-MB-231 cells. ( A ) MDA-MB-231 cells were cultured in glucose-free DMEM without FBS for nutrient starvation (FBS(−) and Glc(−)) or control DMEM with 10% FBS (FBS(+) and Glc(+)) for 12 h before 0–6 Gy irradiation. After irradiation, the cells were cultured under the same condition for 4 h and used for a colony formation assay. The data are presented as the mean ± SD from three independent experiments (*: p < 0.05). ( B , C ) MDA-MB-231 cells were cultured under nutrient starvation for 0–24 h and processed for Western blot analyses with the antibodies indicated. Western blot results at the 0-h time point showed the expression of AMPKα, ATM, DNA-PKcs, FOXO3a, SOD2, Src, EGFR, PDK1, and HIF-1α when the cells were cultured in the control DMEM containing 1.0 g/L glucose with 10% FBS.

Article Snippet: The following antibodies were used as primary antibodies: AMPKα antibody (#2603, Cell Signaling Technology, Inc. (CST), Beverly, MA, USA); phospho-AMPKα (Thr172) antibody (#2535, CST); ATM antibody (#NB100-104, Novus Biologicals, LLC, Englewood, CO, USA); phospho-ATM (Ser1981) antibody (#5883, CST); DNA-PKcs antibody (#sc-9051, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); phospho-DNA-PKcs (Ser2056) antibody (#ab18192, Abcam, PLC, Cambridge, UK); EGFR antibody (#2232, CST); phospho-EGFR (Tyr845) antibody (#2231, CST); FOXO3a antibody (#12829, CST); phospho-FOXO3a (Ser413) antibody (#8174, CST); hypoxia-inducible factor-1 alpha (HIF-1α) antibody (#A300-286A, Bethyl Laboratories, Inc., Montgomery, TX, USA); PDK1 antibody (#3062, CST); phospho-PDK (Ser241) antibody (#3438, CST); SOD2 antibody (#13141, CST); Src antibody (#2108, CST); and phospho-Src (#2102, CST) antibody.

Techniques: Expressing, Cell Culture, Irradiation, Colony Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: AMPK/FOXO3a Pathway Increases Activity and/or Expression of ATM, DNA-PKcs, Src, EGFR, PDK1, and SOD2 and Induces Radioresistance under Nutrient Starvation

doi: 10.3390/ijms241612828

Figure Lengend Snippet: Effects of AMPKα knockdown on phosphorylation and/or expression of ATM, DNA-PKcs, FOXO3a, Src, EGFR, PDK1, and SOD2 in MDA-MB-231 cells. ( A , B ) MDA-MB-231 cells treated with siRNA for AMPKα (siAMPKα) or with control siRNA (siCtrl) were cultured in glucose-free DMEM without FBS for nutrient starvation (FBS(−) and Glc(−)) or in control DMEM with 10% FBS (FBS(+) and Glc(+)) for 12 h and processed for Western blot analyses with the antibodies indicated.

Article Snippet: The following antibodies were used as primary antibodies: AMPKα antibody (#2603, Cell Signaling Technology, Inc. (CST), Beverly, MA, USA); phospho-AMPKα (Thr172) antibody (#2535, CST); ATM antibody (#NB100-104, Novus Biologicals, LLC, Englewood, CO, USA); phospho-ATM (Ser1981) antibody (#5883, CST); DNA-PKcs antibody (#sc-9051, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); phospho-DNA-PKcs (Ser2056) antibody (#ab18192, Abcam, PLC, Cambridge, UK); EGFR antibody (#2232, CST); phospho-EGFR (Tyr845) antibody (#2231, CST); FOXO3a antibody (#12829, CST); phospho-FOXO3a (Ser413) antibody (#8174, CST); hypoxia-inducible factor-1 alpha (HIF-1α) antibody (#A300-286A, Bethyl Laboratories, Inc., Montgomery, TX, USA); PDK1 antibody (#3062, CST); phospho-PDK (Ser241) antibody (#3438, CST); SOD2 antibody (#13141, CST); Src antibody (#2108, CST); and phospho-Src (#2102, CST) antibody.

Techniques: Expressing, Cell Culture, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: AMPK/FOXO3a Pathway Increases Activity and/or Expression of ATM, DNA-PKcs, Src, EGFR, PDK1, and SOD2 and Induces Radioresistance under Nutrient Starvation

doi: 10.3390/ijms241612828

Figure Lengend Snippet: Effects of FOXO3a knockdown on phosphorylation and/or expression of AMPKα, ATM, DNA-PKcs, Src, EGFR, PDK1, and SOD2 in MDA-MB-231 cells. ( A , B ) MDA-MB-231 cells treated with siRNA for FOXO3a (siFOXO3a) or with control siRNA (siCtrl) were cultured in glucose-free DMEM without FBS for nutrient starvation (FBS(−) and Glc(−)) or in control DMEM with 10% FBS (FBS(+) and Glc(+)) for 12 h and processed for Western blot analyses with antibodies indicated.

Article Snippet: The following antibodies were used as primary antibodies: AMPKα antibody (#2603, Cell Signaling Technology, Inc. (CST), Beverly, MA, USA); phospho-AMPKα (Thr172) antibody (#2535, CST); ATM antibody (#NB100-104, Novus Biologicals, LLC, Englewood, CO, USA); phospho-ATM (Ser1981) antibody (#5883, CST); DNA-PKcs antibody (#sc-9051, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); phospho-DNA-PKcs (Ser2056) antibody (#ab18192, Abcam, PLC, Cambridge, UK); EGFR antibody (#2232, CST); phospho-EGFR (Tyr845) antibody (#2231, CST); FOXO3a antibody (#12829, CST); phospho-FOXO3a (Ser413) antibody (#8174, CST); hypoxia-inducible factor-1 alpha (HIF-1α) antibody (#A300-286A, Bethyl Laboratories, Inc., Montgomery, TX, USA); PDK1 antibody (#3062, CST); phospho-PDK (Ser241) antibody (#3438, CST); SOD2 antibody (#13141, CST); Src antibody (#2108, CST); and phospho-Src (#2102, CST) antibody.

Techniques: Expressing, Cell Culture, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: AMPK/FOXO3a Pathway Increases Activity and/or Expression of ATM, DNA-PKcs, Src, EGFR, PDK1, and SOD2 and Induces Radioresistance under Nutrient Starvation

doi: 10.3390/ijms241612828

Figure Lengend Snippet: Suggested molecular pathway for the increased activity and/or expression of AMPK, FOXO3a, ATM, DNA-PKcs, Src, EGFR, PDK1, and SOD2 under nutrient starvation. Our results suggest that nutrient starvation activates the AMPK/FOXO3a pathway and induces radioresistance via increased activity and/or expression of ATM, DNA-PKcs, Src, EGFR, PDK1, and SOD2. AMPK and FOXO3a appear to be key molecules that induce radioresistance under nutrient starvation. Purple arrows indicate activation pathways presented in this manuscript. Red arrows indicate transcriptional activation presented in this manuscript. Black arrows indicate previously reported pathways. Blue arrow indicates transcription.

Article Snippet: The following antibodies were used as primary antibodies: AMPKα antibody (#2603, Cell Signaling Technology, Inc. (CST), Beverly, MA, USA); phospho-AMPKα (Thr172) antibody (#2535, CST); ATM antibody (#NB100-104, Novus Biologicals, LLC, Englewood, CO, USA); phospho-ATM (Ser1981) antibody (#5883, CST); DNA-PKcs antibody (#sc-9051, Santa Cruz Biotechnology, Inc., Dallas, TX, USA); phospho-DNA-PKcs (Ser2056) antibody (#ab18192, Abcam, PLC, Cambridge, UK); EGFR antibody (#2232, CST); phospho-EGFR (Tyr845) antibody (#2231, CST); FOXO3a antibody (#12829, CST); phospho-FOXO3a (Ser413) antibody (#8174, CST); hypoxia-inducible factor-1 alpha (HIF-1α) antibody (#A300-286A, Bethyl Laboratories, Inc., Montgomery, TX, USA); PDK1 antibody (#3062, CST); phospho-PDK (Ser241) antibody (#3438, CST); SOD2 antibody (#13141, CST); Src antibody (#2108, CST); and phospho-Src (#2102, CST) antibody.

Techniques: Activity Assay, Expressing, Activation Assay

Journal: iScience

Article Title: Targeting CD36 determines nicotine derivative NNK-induced lung adenocarcinoma carcinogenesis

doi: 10.1016/j.isci.2023.107477

Figure Lengend Snippet:

Article Snippet: Phospho-Src Family (Tyr416) (D49G4) Rabbit mAb , Cell Signaling Technology , Cat# 6943S; RRID: AB_10013641.

Techniques: Recombinant, Subcloning, Plasmid Preparation, shRNA, Software

Journal: iScience

Article Title: Targeting CD36 determines nicotine derivative NNK-induced lung adenocarcinoma carcinogenesis

doi: 10.1016/j.isci.2023.107477

Figure Lengend Snippet:

Article Snippet: Phospho-Src (Tyr527) Antibody , Cell Signaling Technology , Cat# 2105S; RRID: AB_331034.

Techniques: Recombinant, Subcloning, Plasmid Preparation, shRNA, Software