Fig. 4

A-PRF + stimulates oxidative response, mitochondrial, ER and lysosomal activities in primary equine fibroblasts. (A) Representative images of ROS positive cells after 24 h of treatment. Staining was performed with DCFH-DA. (B) Quantitative analysis of ROS production after a time course of 3, 6 and 24 h. (C) Representative images of TMRE positive cells after 24 h of treatment. (D) Quantitative analysis of TMRE positive cells after 24 h. (E) Representative images of mitochondrial fragmentation process after 24 h of treatment. Staining was performed with mitotracker (red, mitochondria) and Hoechst 33,342 (blue, nuclei). (F) Quantitative analysis of mitochondrial length. (G) Representative images of AO staining after 24 h of treatment. (H) Quantitative analysis of AO cells after 24 h. (I) Representative images of ER expansion after 24 h of treatment. Staining was performed with ER-ID (red, ER) and Hoechst 33,342 (blue, nuclei). (J) Quantitative analysis of ER expansion after 24 h. Scale bars (Figs. 4A, 4C, 4G, 4I): 100 μm. N ≥ 3. Scale bars (Fig. 4E): 2.5 μm. N ≥ 10. Results are expressed as mean ± standard deviation (SD) of three independent experiments. # denotes p < 0.05 versus FBS 20%; *, **, *** denote respectively p < 0.05, p < 0.01 and p < 0.001 versus Ctrl.