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. 2019 May 2;142(4):328–350. doi: 10.1159/000499506

Fig. 6.

Fig. 6

Clinically proven anti-fibrotics, Nintedanib and Fresolumimab, effectively inhibit ECM accumulation. a Effect of a marketed anti-fibrotic, Nintedanib, on ECM in the co-culture of epithelial cells and fibroblasts. Cells were cultured for 7 days in co-culture in the presence of Nintedanib (10–0.02 µM). Cells were then lysed, and the ECM was fixed and stained using a total protein dye or specific antibodies. i Graph shows the effect of Nintedanib on total ECM components (total protein dye, solid line) or on cell viability (PrestoBlue, dashed line). Data are represented as log2 fold change from the co-culture in the presence of 0.1% DMSO and shows mean ± SD from a representative of 3 independent experiments with 4 replicate wells per condition. IC50s estimated for the total and individual ECM markers from the independent experiments are presented as geomeans with the range in Table 2. ii Corresponding images of total ECM components (total protein dye, white) for 5 µM Nintedanib or DMSO control (0.1% DMSO). b Effect of an anti-TGFB antibody on ECM accumulation in the co-culture of epithelial cells and fibroblasts. Cells were cultured for 7 days in the presence of anti-TGFB antibody (MAB1835) or isotype control (MAB002; 4,000–0.12 nM). Cells were then lysed, and the ECM was fixed and stained using a total protein dye or specific antibodies. i Graph shows the effect of the anti-TGFB antibody (in blue) on total ECM. Corresponding isotype control is also shown (in black). There was no effect on cell viability (PrestoBlue) at any concentration tested. Data are represented as log2 fold change from the untreated co-culture and show mean ± SD from a representative of 4 independent experiments with 4 replicate wells per condition. IC50s estimated for the total and individual ECM markers from the independent experiments are summarised as geomeans with the range in Table 3. ii Corresponding images of total ECM components (total protein dye, white) for 7.8 or 1,000 nM anti-TGFB antibody or isotope control. Images show a single field. Scale bars represent 100 µm. ECM, extracellular matrix.