Ab8129

Paraffin-embedded subcutaneous tumor tissues were sectioned (5 μM thick) and immunostained with pSTAT3, proliferating cell nuclear antigen (PCNA), Iba-1, CD3, CD34, and CD204. Anti-pSTAT3 (D3A7, Cell Signaling Technology; 1:200), anti-PCNA (M0879, DAKO; 1:200), anti-Iba-1 (019-19741, Wako; 1:4000), anti-CD3 (413591, Nichirei Biosciences; 1:2), anti-CD34 (ab8129, Abcam; 1:2000), and anti-CD204 (2F8, Invitrogen; 1:2000) antibodies were used as primary antibodies. The sections were subsequently treated with an HRP-conjugated secondary antibody (Nichirei, Tokyo, Japan), followed by the visualization with diaminobenzidine.

The following antibodies were used in this study: anti-FLAG (WB 1:1,000, F3165; Sigma), anti-EPLIN (WB 1:1,000, IF 1:100, 50311; Cell Signaling Technology), anti-EPLIN (WB 1:1,000, IF 1:100, sc-136399; Santa Cruz Biotechnology), anti-EPLIN (WB 1:1,000, immunoprecipitation [IP] 1 μg/1 mg lysate, immunohistochemistry [IHC] 1:200, 16639–1-AP; Proteintech), anti-HA (WB 1:500, IP 2 µg/1 mg cell lysate, SC F-7; Santa Cruz Biotechnology), Alexa Fluor 568–phalloidin (IF 1:1,000, A1238; Life Technologies), SiR-actin (1:10,000, CY-SC001; Cytoskeleton), anti–β-tubulin (WB 1:2,500, 926–42211; LI-COR), anti–α-tubulin (WB 1:2,500, 23948; Santa Cruz Biotechnology), pFAK Y397 (WB 1:1,000, ab8129; Abcam), FAK S910 (WB 1:1,000, 44-596G; Invitrogen), anti–E-Cadherin (WB 1:1,000, IHC 1:200, 3195; Cell Signaling Technology), total FAK (WB 1:1,000, 610087; BD Biosciences), anti-zyxin (IF 1:1,000, ab50391; Abcam), anti-Rab40b (WB 1:500, LS-C353287; LSBio), anti-Rab40c (WB 1:500, H-8 sc-514826; Santa Cruz Biotechnology), and anti-CD63 (IF 1:100; gift from Andrew Peden, University of Shefield, Shefield, UK).