Figure 1

TGFβ1 stimulates the expression of ECM components in human and rabbit primary cells. (A) Selected ECM mRNA transcripts were measured using qRT-PCR in human RPTEC or human IPF134 following 48 hr stimulation with TGFβ1 (10 ng/mL). Data show transcript abundance relative to the unstimulated control (indicated by the dashed line) and are shown as mean ± SD of four (RPTEC) or seven (IPF134) independent experiments. ****P < 0.0001; ****P < 0.001; **P < 0.01; Student’s t-test. (B) The accumulation of ECM in human RPTEC (i) or human IPF (ii) cells stimulated with 10ng/mL TGFβ1 was evaluated via measurement of the incorporation of ¹⁴C-labelled amino acids into the deposited ECM. Data presented are the mean ± SD of three independent experiments. **P < 0.01; Student’s t-test. (C) Increased mature ECM accumulation following treatment of cells with a pro-fibrotic stimulus can be visualised following in-situ fluorescent staining using the Flamingo dye. Cells from three different systems (primary human RPTEC; primary human IPF134; co-culture of primary rabbit RPTEC with primary rabbit renal fibroblasts) were cultured in the absence (top) or presence (bottom) of 10 ng/ml TGFβ1 before decellularised matrix was fixed and stained in situ with Flamingo fluorescent dye. Images show a single field.