Fig. 3.

Effects of ADAM10 shRNA depletion in mesothelioma cells on proliferation, migration and invasion. a Western blot analysis of ADAM10 protein levels in AB12 and PM27 cells transduced with two different control shRNA (shCtl[1] or [2]) or with a shRNA targeting ADAM10 (shADAM10). Actin was used as a loading control. b Proliferation rates were assessed for AB12 and PM27 cells after 4, 24, 48 and 72 h by CYQUANT analysis (n ≥ 6). All measurements were normalized to the ‘4 h-timing’ considered as baseline proliferation and expressed as percentage increase from baseline. For in vitro results, shCtl condition correspond to shCtl[1]-treated AB12 or PM27 cells. c Representative examples of AB12 and PM27 percentages present in the different phases of cell cycle (G1, S or G2) 24 h after cell seeding. d Upper panel: representative HE-stained images showing cells that migrated through the transwell filter. Scale bars = 100 µm. Lower panel: quantification of numbers of migrating AB12 or PM27 cells transduced with shCtl or shADAM10 per field (n = 3). Results are expressed as percentage of control condition (mean ± SEM); Student t test; **P < 0.01. e Wound healing assay performed on AB12 and PM27 cells transduced with shCtl or shADAM10. The wound closure was evaluated 4, 6 and 8 h after the scratch (n = 8). Results are expressed as mean ± SEM; Student t test; *P < 0.05; ***P < 0.001. f Upper panel: representative examples of spheroids composed of AB12 or PM27 cells transduced with shCtl or shADAM10; magnification ×10. Lower panel: quantification of cell invasion in collagen gel (n ≥ 7). Results are expressed as mean ± SEM; Student t test; *P < 0.05; ***P < 0.001