Journal: Cell Reports Medicine

Article Title: Lung adenocarcinoma surfaceome remodeling with EGFR inhibitors uncovers placental alkaline phosphatase as a target for combination therapy

doi: 10.1016/j.xcrm.2025.102513

Figure Lengend Snippet: EGFR TKI dephosphorylates FoxO3a to induce ALPP expression (A and B) Immunoblots for ALPP, pAKT-S473, total AKT, pERK-T202/Y204, total ERK, pSTAT1-Y701, pSTAT1-S727, total STAT1, and β-actin in LUAD cancer cells H2291 and H1650 following 24- and 48-h treatment with or without EGF (10 or 20 ng/mL) or gefitinib (0.5 or 1 μM). Representative blot of three replicates. (C) Immunoblots for ALPP and β-actin in HCC827 and PC9 cell treated with PI3K inhibitor NVP-BKM120 (100 nM), MEK inhibitor AZD8330 (100 nM), and ERK inhibitor SCH772984 (100 nM) for 48 h. Representative blot of three replicates. (D and E) Immunoblots for ALPP and β-actin in HCC827 and PC9 cell treated with FOXO inhibitor AS1842856 with vehicle or gefitinib for 48 h. Representative blot of three replicates. (F) Immunoblots for ALPP and FoxO3a in FoxO3a-overexpressing HCC827 and H1650 LUAD cells treated with vehicle or gefitinib (30 nM) for 48 h. Representative blot of three replicates. (G) Immunoblots for ALPP, FoxO3a, and phosphorylated FoxO3a (Ser294 and Ser425) in HCC827 and H1650 cells treated with 30 nM gefitinib for 6 h. Representative blot of three replicates. (H) Immunoblots for FoxO3a in the cytosol and nucleus compartment from gefitinib-treated HC827 and H1650 cell lines. Representative blot of three replicates. (I) Immunoblots for ALPP, EGFR, pEGFR (Tyr1068), FoxO3a, pFoxO3a (Ser294), and β-Actin in HCC827 and PC9 cells treated with gefitinib (30 nM) or osimertinib (30 nM). Representative blot of three replicates. (J) ChIP-qPCR assay for the promoter region of ALPP gene in HCC827 and H1650 LUAD cells treated with either vehicle or gefitinib (1 μM) for 6 h ( n = 3 biological replicates). Data are presented as mean ± SD. Statistical significance was determined using Dunnett’s multiple comparisons test, and adjusted p values are reported.

Article Snippet: FoxO inhibitor , MedChemExpress , Cat# AS1842856.

Techniques: Expressing, Western Blot, ChIP-qPCR