Fig 2: CXADR expression in various cell lines and development of anti-CXADR antibodies.(a) Expression of CXADR and tubulin in the indicated cell lines was detected by Western blotting. The gels have been run under the same experimental conditions and cropped to show protein bands corresponding to CXADR or tubulin as indicated. (b) Ba/F3 cells expressing human CXADR were incubated with the indicated antibody clones (gray) or isotype control antibodies (white), followed by by flow cytometry. (c) The positions in CXADR that are bound by the indicated antibody clones are shown.

Fig 3: Effect of anti-CXADR antibody 6G10A on ADCC and CDC.(a) For ADCC activity, DU-145 cells were incubated with splenocytes from male nude mice in the presence of the indicated antibodies at 100 µg/ml for 4 h. For CDC activity, DU-145 cells were incubated with 10% rabbit complement in the presence of the indicated antibodies for 4 h. Cell lysis was determined using calcein AM. The values are means ± SEM (n = 3). *P < 0.05 and **P < 0.01 versus the control values. (b) DU-145 cells were injected subcutaneously into male nude mice. To deplete NK cells in the mice, anti-asialo GM1 antibodies (100 µg/day) were administered intravenously -1, 6, and 13 days after the cancer cell injection. Anti-CXADR antibody 6G10A (250 µg/day) was administered intravenously 1, 7, and 14 days after the cancer cell injection. The mice were sacrificed 21 days after the cancer cell injection, and the tumors were excised and weighed. The values are means ± SEM (n = 5). *P < 0.05 and **P < 0.01 versus the control values.

Fig 4: Effect of anti-CXADR antibody 6G10A on the growth of various subcutaneous tumors in vivo.DU-145, BxPC-3, or DLD-1 cells were injected subcutaneously into male nude mice. Antibodies (250 µg/day) were administered intravenously 1, 7, and 14 days after the cancer cell injection. Mice were sacrificed 21 days after the cancer cell injection, and the tumors were excised and weighed. The values are means ± SEM (n = 3). *P,0.05 and **P < 0.01 versus the control values.

Fig 5: CXADR expression in human normal and tumor tissues.(a) CXADR expression in the indicated tissues was determined by Western blotting using lysates from various tissues. DU-145 cells were used as a positive control. N, normal; T, tumor. The gels have been run under the same experimental conditions and cropped to show protein bands corresponding to CXADR or GAPDH as indicated. (b) Frozen sections from the indicated tumor tissues (BioChain) were stained with anti-CXADR antibody 6G10A. Scale bar is 100 µm.