Journal: ImmunoTargets and Therapy
Article Title: Kasugamycin Inhibits Melanoma Lung Metastasis and CHI3L1-Driven M2-Like Tumor-Associated Macrophage Differentiation
doi: 10.2147/ITT.S563951
Figure Lengend Snippet: KSM inhibits CHI3L1-stimulated Erk and Akt activation and the antiapoptotic effect of CHI3L1 in macrophages. ( A ) AMJ-C11 mouse lung macrophages were stimulated with recombinant (r) CHI3L1 overnight, and the activation of Erk and Akt was detected by Western blot evaluations. pErk, phosphorylated Erk; T-Erk, total Erk; pAkt, phosphorylated Akt, T-Akt, total Akt. Right panel, densitometric quantitation on the blots of pErk and pAkt. ( B ) After exposure of AMJ-C11 cells with H 2 O 2 (1 mM, 24 hours) with and without rCHI3L1 and KSM treatment, cellular apoptosis (% of Annexin V+ cells) was measured by FACS evaluations. ( A and B ) are representatives of 3 independent experiments. Right panel, the values of Annexin 5 (+) cells (%) are the mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001 (One-Way ANOVA, multiple comparisons).
Article Snippet: Protein lysates (25 μg) from lung tissues or cells were subjected to SDS-PAGE, transferred to membranes, and immunoblotted with primary antibodies against CD163 (#PA5-109327, Thermo Fisher Scientific), CD206 (E6T5J, #24595S, Cell Signaling Technology), phosphorylated AKT (p-Akt) (193H12, #4058S, Cell Signaling Technology), and total Akt (11E7, #4685S, Cell Signaling Technology), phosphorylated EGFR (Tyr1068) (p-EGFR) (#44-788G, Thermo Fisher Scientific), EGFR (D38B1, #4267S, Cell Signaling Technology), phosphorylated Erk (p-Erk) (#9101S, Cell Signaling Technology), total Erk (#9102S, Cell Signaling Technology), phosphorylated STAT3 (Tyr705) (p-STAT3) (D3A7, #9145, Cell Signaling Technology), total STAT3 (79D7, #4904, Cell Signaling Technology) and b-actin (C4, #sc-47778 HRP, Santa Cruz Biotechnology).
Techniques: Activation Assay, Recombinant, Western Blot, Quantitation Assay