Journal: Molecular Cancer Research

Article Title: β8 Integrin Mediates Pancreatic Cancer Cell Radiochemoresistance

doi: 10.1158/1541-7786.mcr-18-1352

Figure Lengend Snippet: Figure 5. b8 integrin interactome and subcellular colocalization. A, Sequential immunoprecipitation-mass spectrometry–based analysis of b8 integrin interactome was performed in unirradiated and 6-Gy X-rays irradiated Patu8902 cells. Time point of analysis after irradiation was 2 hours. B, Comparative changes in the b8 integrin interactome between control and irradiation upon categorization into different molecular functions using the PANTHER classification system. C, Immunofluorescence costaining of b8 integrin with GM130 (Golgi), mitochondria, aV integrin, APPL2, and Caveolin 1. Scale bars, 10 mm. D, Pearson correlation analysis of C. Data show mean SD (n ¼ 3).

Article Snippet: The antibodies against b1 integrin (WB 1:1,000, Abcam, ab179471), b8 integrin (IF 1:200, WB 1:1,000; Abcam, ab80673, Rb, recognizes aa 614-663 (this antibody was generally used in this study); Abnova, H00003696-M01, Mo, recognizes aa 392–503 (this antibody was used only where specifically indicated), GM130 (IF 1:100, WB 1:1,000, BD, 610822), MEK1/2 (WB 1:1,000, Cell Signaling Technology, 4694s), gH2AX (WB 1:1,000, Cell Signaling Technology, 9718s), aV integrin (IF 1:250, Novus, NB100-2618), APPL2 (IF 1:100, Sigma-Aldrich, SAB1400605), Caveolin 1 (IF 1:100, BD, 610407), LC3B (IF 1:500, Sigma-Aldrich, SAB4200361), b-actin (WB 1:10,000, Sigma-Aldrich, A5441), HRP-conjugated secondary antibody (GE, Rb NXA931, Mo NXA931), Alexa Fluor 488– or Alexa Fluor 594–conjugated secondary antibodies (1:500, Life Technologies) were purchased and used as indicated.

Techniques: Immunoprecipitation, Mass Spectrometry, Irradiation, Control