Figure 7

(A) Schematic image of co-culture. XL-2 cells, human hepatic stellate cell line, were co-cultured with primary human macrophages (MΦs) using a filter for 48 h in the presence or absence of lipopolysaccharides (LPS), recombinant human transforming growth factor beta 1 (TGFβ1) (active form) and/or latency associated peptide (LAP); subsequently, the underlying XL-2 cells were subjected to the following analysis. (B) Phase contrast images of macrophages cultured in the upper layer with or without LPS-administration alone. (C) Immunocytochemical staining images. Forty-eight after XL-2 cells were co-cultured with macrophages under the indicated each condition, immunocytochemical staining was performed using anti-collagen type 1a (COL1A1) antibody. 3,3′-Diaminobenzidine (DAB) was used to detect the COL1A1 signals (brown). Nuclei stained with hematoxylin (blue). (D) Expressions of COL1A1 and the inflammation-related genes. Real-time RT-PCR was performed to investigate the mRNA expression level of the indicated genes in the cells cultured under the same conditions as in (B). Quantitative data were presented as the mean ± SEM (n = 5). The letters above each bar in the graph indicate statistically significant differences (P < 0.05) compared to the corresponding sample group.