Extended Data Fig. 8: Secreted IGFBP6 and MVP do not affect inflammation in HUVECs.
From: Endothelial IGFBP6 suppresses vascular inflammation and atherosclerosis
a and b, HUVECs were treated with adenoviruses (Ad-MVP and Ad-IGFBP6) for 7 days, during which the supernatant was continuously collected. Ultrafiltration tubes were used to concentrate the protein in the supernatant, and flag (a, n = 3) or HA (b, n = 3) antibodies were used for Co-IP, respectively, and finally verified by Western blot. c, HUVECs were treated with different concentrations of IGFBP6 recombinant protein for 42 h, and then treated with TNF-α for 6 h, and indicated proteins were detected by Western blot (n = 3). d, HUVECs were treated with different concentrations of IGFBP6 recombinant protein for 3 h, and then treated with TNF-α for 10 mins, and indicated proteins were detected by Western blot (n = 3). e, 80% of confluent HUVECs were treated with Ad-NC or Ad-IGFBP6 in the presence of TNF. After that, whole cell lysate was collected for Western blot (left 4 lanes, control). In parallel experiments, the treated cells were washed with PBS and new complete medium was supplemented. Then, conditioned media were collected to treat HUVECs before lysate was collected for Western blot (right 4 lanes) (n = 3). f, HUVECs were treated with different concentrations of MVP recombinant protein for 3 h, and then treated with TNF-α for 10 mins, and indicated proteins were detected by Western blot (n = 3).
