Figure 8. Upregulation of H19 lncRNA in ischaemic hindlimbs treated with PGI₂-hMSCs.

(a) Representative images of H19 RNA fluorescence in situ hybridization in gastrocnemius muscle sections at 3 days after 3.1-hMSC, 3.1-hMSC+ILO or PG₂-hMSC injections. We used a fluorescein-tagged H19 FISH probe that specifically targets endogenous H19 lncRNA, resulting in intense intracellular green fluorescent particles. We found a higher expression of host H19 lncRNA in PGI₂-hMSC- and in 3.1-hMSC+ILO-treated muscles than in 3.1-hMSC-treated muscles (n=3 mice per group). All the sections were counterstained with 4,6-diamidino-2-phenylindole (DAPI) to localize nuclei. (b) Quantitative RT–PCR showed a significant increase in H19 lncRNA levels in ischaemic gastrocnemius muscle 5 days after injections with 3.1-hMSCs+ILO or PG₂-hMSCs as compared with tissue samples injected with 3.1-hMSCs. *P<0.05 by one-way ANOVA with Newman–Keuls post hoc test. Data are shown as mean±s.e.m. from three independent assays. N=3 mice (male and female) per group. Mice were sex matched among the groups. Scale bar, 10 μm.