Journal: bioRxiv

Article Title: RhoA/ROCK2 signalling is enhanced by PDGF-AA in fibro-adipogenic progenitor cells in DMD

doi: 10.1101/2021.04.12.439417

Figure Lengend Snippet: A) Representative images of the DMD cells. CD56-were stained with desmin and TE-7 markers. B) Analysis of the PDGFRα in the CD56-isolated cells showing positivity for the receptor. C) Experimental plan to screen. D) Representative WB of total protein bands measured with the Revert 700. E) Representative in-cell western image showing the total cell number signal stained with TO-PRO-3. F) Viability determination using the PrestoBlue cell viability assay to compare the effect of PDGF-AA, C3 and fasudil on FAPs at 30min, 24h, 48h and 72h.Data are represented as the mean of 3 replicates ± standard error of the mean. Results were statistically analysed using one-way ANOVA followed by Tukey post hoc test.

Article Snippet: The negative fraction (CD56-) was cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) and 199 (M199) medium (3: 1) supplemented with 10% fetal bovine serum (FBS),10 mM glutamine and penicillin-streptomycin (all from Lonza, Basel, Switzerland) and their purity was verified with the fibroblast marker TE-7 (Merck) by immunofluorescence (IF) and PDGFRα expression by fow cytometry.

Techniques: Staining, Isolation, In-Cell ELISA, Viability Assay