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. 2022 Feb 28;17(10):2238–2246. doi: 10.4103/1673-5374.336871

Figure 5.

Figure 5

MiR-21a-5p in MSCs-EVs targets STAT3 signaling pathway after OGD in BV-2 cells.

(A) Western blot analysis of phosphorylated STAT3 (p-STAT3) and STAT3 in BV-2 cells after 1-, 3-, and 5-hour OGD followed by 24-hour reoxygenation. The control cells were cultured normally without OGD. (B) Western blot analysis of the levels of p-STAT3 and STAT3 in the presence or absence of MSCs-EVs. The BV-2 cells had undergone 3-hour OGD followed by 24-hour reoxygenation. (C) Western blot analysis of the levels of p-STAT3 and STAT3 in BV-2 cells treated with EVs, EVs-miR-21ainhibitor, or EVs-miR-21aINC. The cells had undergone 3-hour OGD followed by 24-hour reoxygenation. (D) The sequence of mouse miR-21a-5p and its predicted binding site within the STAT3 3’ untranslated region (3′-UTR); the mutated sequence is also shown. (E) Luciferase activity in BV-2 cells transfected with luciferase plasmids containing WT STAT3 3′-UTR or MUT STAT3 3′-UTR and treated with miR-21a-5p mimics or miR-21a-5p NC. The cells were lysed to measure the relative luciferase activity. The experiments were carried out on four independent samples. Graphs show the mean ± SD; *P < 0.05, **P < 0.01, ***P < 0.001 (independent samples t-test) in A and E; *P < 0.05 (one-way analysis of variance with Bonferroni correction) in B and C. EV: Extracellular vesicle; MSC: mesenchymal stromal cell; OGD oxygen-glucose deprivation.