Fig. 2.

Migration, proliferation, viability, and activated signaling pathways in HSCs exposed to CM from MerTK-inhibited THP-1 macrophages.

(A-C-E) Serum-starved HSCs were exposed to SF medium (Control, white columns) or to CM from THP-1 macrophages left untreated (DMSO, purple columns), treated with UNC569 (light purple columns) or stimulated with Gas-6 in the presence (light purple columns) or absence (purple columns) of UNC569. (B-D-F) Serum-starved HSCs were exposed to SF medium (Control, white columns) or to CM of THP-1 macrophages transfected with NT siRNA (purple columns) or MERTK-siRNA (light purple columns) and left untreated or treated with Gas-6. (A-B) HSC migration was measured in modified Boyden chambers. (C-D) HSC proliferation was assayed by BrdU Cell Proliferation ELISA Kit. (E-F) HSC viability was measured by MTT assay. (G) Serum-starved HSCs were exposed to CM from THP-1 macrophages unstimulated or stimulated Gas-6 in the presence or absence of UNC569 or (H) to CM from THP-1 macrophages transfected with NT siRNA or MERTK-siRNA stimulated with or without Gas-6 for 24 h. 30 μg of total cell lysates were subjected to immunoblot analysis to detect different proteins or phosphoproteins. Densitometries of p-STAT3/STAT3, p-P38/P38 and IL8/β-Actin expression (n = 3) were shown in the graphs. Data are mean ± SEM. Statistical significance was assessed by Student's t test. ∗p <0.05 vs. SF medium; ∗∗p <0.05 vs. CM of untreated THP-1 macrophages; ^§p <0.05 vs. CM of THP-1 macrophage stimulated with Gas-6 without inhibition for MerTK; ^†p <0.05 vs. SF medium; ^††p <0.05 vs. CM of NT siRNA unstimulated THP-1 macrophages; ^¥p <0.05 vs. CM of NT siRNA THP-1 macrophage stimulated with Gas-6. CM, conditioned media; HSC(s), hepatic stellate cells; NT, non-targeting; SF, serum free; siRNA, small-interfering RNA.