Skip to main content
. 2020 May 25;11:2604. doi: 10.1038/s41467-020-16409-z

Fig. 5. Activin regulates pro-fibrotic ECM deposition by fibroblasts.

Fig. 5

a–d HaCaT keratinocytes transduced with lentiviruses allowing Dox-inducible overexpression of INHBA (ActOE) or empty vector (EV) were co-cultured with murine immortalized fibroblasts (GFP-expressing) transduced with lentiviruses allowing Dox-inducible expression of dnActRIB or with EV-transduced fibroblasts for 7 days in 1% or 10% FBS. Co-cultures were stained for collagen type I (a, b), periostin, and asporin (c, d). The experiment was repeated twice with similar results. Representative photomicrographs of ECM staining for each group. Bar graphs show quantification of the percentage of the stained area on whole co-culture coverslips. Scale bars: 1000 μm. n = 3. e, f Representative high magnification photomicrographs of collagen type I (e) and asporin (f) staining of ActOE/EV and ActOE/dnActRIB co-cultures in the presence of 1% or 10% FBS. GFP-positive fibroblasts are shown (green); nuclei were counterstained with DAPI (blue). Scale bars: 100 μm. g, h GFP-expressing fibroblasts were sorted from GFP-negative HaCaT keratinocytes after 8 days co-culture, and gene expression was analyzed by qRT-PCR relative to RPL27 for keratinocytes (g) or Rps29 for fibroblasts (h). n = 3. The experiment was repeated twice with similar results. Graphs show mean ± SEM and P values; mean expression levels in AA cultures were set to 1 in g and h; one-way ANOVA and Tukey’s multiple comparison post hoc tests (a–d, g, h). All n numbers indicate biological replicates. AA, EV/EV; BA, ActOE/EV; AB, EV/dnActRIB; BB, ActOE/dnActRIB. Source data are provided as a Source Data file (Fig. 5).