Fig. 4. Plasminogen treatment regulates the expression of genes responsible for the formation and remodeling of granulation tissue.

a The KEGG enrichment scatter plot of genes involved in the formation and remodeling of granulation tissue that were significantly downregulated in plasminogen-treated wounds at day 20 when compared with PBS-treated wounds, as identified using mRNA sequencing. b, c Expression of Actc1 and Tgfb1 (respectively) in radiation wounds before treatment (day 0) (n = 3) and at different time points of PBS (black) (n = 3) or plasminogen (red) (n = 3) treatment as measured with the Mouse Wound Healing RT² Profiler PCR array. The mRNA levels for all samples are presented as fold regulation relative to mRNA levels in the control healthy, nonirradiated skin. d The expression of genes involved in the TGF-β pathway that were significantly changed in plasminogen-treated wounds at day 20 when compared with PBS-treated wounds, as measured using mRNA sequencing. e, f Quantification of CTGF and TGF-β (respectively) protein levels in healthy non-radiated skin, in irradiated skin before treatment (day 0), and in PBS- and plasminogen-treated radiation wounds at days 20 and 30, based on immunostaining of paraffin sections from the wounded area (n ≥ 3). g Expression of genes involved in the WNT pathway measured as described in panel d. h Expression of Wisp1 in radiation wounds shown as described in panel b. i Quantification of FOSL1 protein levels as described in panel e (n ≥ 3). j Fold regulation of MAPK pathway-related genes that were significantly enriched at day 20 of plasminogen- compared with PBS treatment. k Quantification of CD14 protein levels as described in panel e (n ≥ 3). l Expression of Hgf in radiation wounds measured as described in panel b. m Quantification of Ki67 protein levels as described in panel e (n ≥ 3). n, o Expression of Timp1 and Serpine1 in radiation wounds, respectively, shown as described in panel b. p Quantification of CD31 protein levels as described in panel e (n ≥ 3). q, r Expression of Itga5 and Mmp9 (respectively) in radiation wounds, shown as described in panel b. *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001. Rich factor is the ratio of the number of target genes divided by the number of all the genes in each pathway.