Figure 3
IL-8 is required for the growth factor-like activity of TpF1.
(a) HUVECs were exposed to TpF1, as in Fig. 1a, in the presence or absence of 1 μg/ml anti-IL-8 blocking antibody. After 12 h cells were counted and normalized to the number of plated cells set as 1 A.U. The graph shows cell proliferation under the different conditions. Data are represented as the mean ± S.D. of three independent experiments. (b) HUVECs, grown to confluence in 6-well plates, were exposed to TpF1 for 24 h. Supernatants were collected and incubated or not with anti-IL-8 blocking antibody for 45 min, before transferring to the lower chamber of 24 Transwell plates with Fast DiI-labelled HUVECs seeded on the upper chamber. Migrated cells were quantified after 2 and 6 h. Data are represented as the mean ± S.D. of three independent experiments. (c) HUVECs seeded on Matrigel-coated coverslips were exposed to TpF1 for 12 h, in the presence or absence of the IL-8 blocking antibody. Cells were stained with Phalloidin-Alexa Fluor 546 and analyzed by confocal microscopy at 200 × magnification. Scale bar = 300 μm.
