Mice were given intraperitoneal (i.p.) injections of 2 μl per g body weight CCl4 (Sigma) at ratio of 1:3 by volume in olive oil (Sigma) or olive oil alone (control) twice weekly for 8 weeks.
For the VP (VP) experiments, fibrosis was induced by CCl4 injections for 6 weeks with VP treatment for the final 3 weeks. VP was dissolved in 10% DMSO in PBS to a concentration of 10 mg ml−1. Ten microlitre per g body weight (100 mg VP/kg bodyweight) was injected i.p. three times per week (on alternate days to CCl4 or olive oil). Control mice were injected with DMSO. This created four treatment groups: olive oil+DMSO (n=5); olive oil+VP (n=3); CCl4+DMSO (n=3); and CCl4+VP (n=4).
Treatment with IPA3 (Tocris Bioscience) or DMSO (Sigma) occurred during the final 4 weeks. IPA3 was dissolved in DMSO to a concentration of 4 μg μl−1; 1 μl per g body weight (4 mg IPA3/kg bodyweight) was injected i.p. three times per week (on alternate days to CCl4 or olive oil injections). Control mice were injected with DMSO. This created four treatment groups: olive oil+DMSO (n=5); olive oil+IPA3 (n=5); CCl4+DMSO (n=5); and CCl4+IPA3 (n=5).
Martin K., Pritchett J., Llewellyn J., Mullan A.F., Athwal V.S., Dobie R., Harvey E., Zeef L., Farrow S., Streuli C., Henderson N.C., Friedman S.L., Hanley N.A, & Piper Hanley K. (2016). PAK proteins and YAP-1 signalling downstream of integrin beta-1 in myofibroblasts promote liver fibrosis. Nature Communications, 7, 12502.
+ Open protocol