HCMV secretome stimulates survivin expression through IL-6 receptor blocking apoptosis and promoting AG. (A) Cell lysates from HUVECs incubated with mock-secretome (MS) and HCMV VR1814 (VRS) for 24, 48, 72, and 96 hours were subjected to SDS-PAGE followed by Western blotting to evaluate the expression of survivin. αTubulin served as an internal control. (B) HUVECs were treated with anti–IL-6R antibody and stimulated with the secretome for 48 hours. Expression of survivin, cleaved caspase-3, and caspase-7 was determined by Western blotting. αTubulin immunodetected with mAb served as an internal control. (C) HUVECs were infected with recombinant retroviruses expressing either a survivin-directed shRNA or a nonspecific control (scramble) shRNA followed by stimulation with the HCMV secretome. Expression of STAT-3, survivin, cleaved caspase-3, and caspase-7 was determined by Western blotting. αTubulin served as an internal control. (D) Quantification of Matrigel tube formation assay after 24 hours of HUVECs in the same experimental condition as above. The results are expressed as mean ± SD, ^###P < .01 versus mock. (B) Representative examples of each culture condition are shown as a low-power image (magnification, ×10).