Figure 2.

TGF-β1 treatment of 3D human liver spheroids increases fibrotic markers as well as CTGF. Three-dimensional human liver spheroids were treated with 5 ng/mL of TGF-β1 for 3 days. Blue bars represent mRNA expression data. Red bars show results of IHC analysis, where each dot represents an average result of at least 10 analyzed sections of spheroids in 1 separate experiment. Lastly, green bars represent results of ELISA analyses. (A–C,G,H) Transcription of COL1A1, TGFB1, LOX and VIM was increased 3 days after treatment initiation, while ACTA2 expression was unaffected (n = 3). (D–F) Representative images of key fibrosis related markers at day 10 including quantification of the increase of COL1A1, TGF-β1 and α-SMA expression are shown (n = 3). (I) TGF-β1 treatment increased the pro-collagen I α1 and CTGF released into the culture medium. (J) CTGF was increased in both monoculture (left) and co-culture (middle) spheroids in response to TGF-β1 treatment (n = 3), which is also visualized by immunofluorescence (right). In the Figure 2, all of the data presented, with the exception of (J) ((left), monocultures), represents experiments performed in co-culture spheroids. Nuclei in the IHC images are shown in blue. The size of the spheroids is marked with a white scale bar of 100 μm. Data are shown as mean ± SEM. Statistical analysis—Student’s t-test; * p < 0.05; ** p < 0.01; ns: not significant.