src family antibody sampler kit  (Cell Signaling Technology Inc)

Journal: Frontiers in immunology

Article Title: Tumor Cell-Secreted ISG15 Promotes Tumor Cell Migration and Immune Suppression by Inducing the Macrophage M2-Like Phenotype.

doi: 10.3389/fimmu.2020.594775

Figure Lengend Snippet: FIGURE 4 | Interferon-stimulated gene 15 (ISG15)-induced CCL18 secretion by macrophages was dependent on the ISG15 receptor, leukocyte function-associated antigen-1 (LFA-1), and SRC family kinase (SFK) signaling. (A) Immunofluorescence demonstrated that the ISG15 receptor CD11a/CD18, also called LFA-1, was expressed on the membranes of human macrophages. Macrophages were treated with 295nM rISG15 for 24 h. Scale bars, 10 mm. (B) ELISA results showed small molecular inhibitors of LFA-1, A286982, inhibited CCL18 secretion by macrophages. (C) Small molecular inhibitors of LFA-1 and SRC, A286982 and PP2, hindered the ISG15-induced activation of SFK signaling. (D) PP2 reduced the CCL18 secretion by ISG15-treated macrophages in a dose-dependent manner, as determined by ELISA. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Protein extracts were resolved by 10% SDS–PAGE, transferred to PVDF membranes (Roche), and probed with antibodies directed Frontiers in Immunology | www.frontiersin.org 4 against human ISG15 (1:1,000; Abnova, catalog no. A155801), SRC (1:1,000; CST, catalog no. 9935T), and b-actin (1:3,000; Abcam, catalog no. ab69512).

Techniques: Enzyme-linked Immunosorbent Assay, Activation Assay