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. 2021 Mar 25;11:618662. doi: 10.3389/fonc.2021.618662

Figure 4.

Figure 4

miR-19a-3p enhances osteoclastogenesis and activation through PI3K/AKT signaling pathway. (A) Transfection efficiency of miR-19a-3p mimics in RAW264.7 cells. miR-19a-3p expression was analyzed by qRT-PCR after 24 hours post transfection with mimics. (B) Representative images showing the osteoclastogenesis, podosome belts and hydroxyapatite resorption in each group and corresponding statistics. (C, D) Representative qRT-PCR and Western Blot analysis of miR-19a-3p, PTEN, MMP-9, CTSK, p-AKT and AKT expression. RAW264.7 cells transfected with miR-19a-3p mimic-NC or mimics were stimulated with RANKL (50 ng/mL) for 4 days before RNA and protein collection. (E) QRT-PCR analysis of miR-19a-3p level in RAW264.7 cells after 24 hours post transfection with inhibitor-NC or inhibitor. (F) Representative images showing the osteoclastogenesis, podosome belts and hydroxyapatite resorption in each group and corresponding statistics. (G, H) Representative qRT-PCR and Western Blot results of the effects of miR-19a-3p inhibitor on miR-19a-3p, PTEN, MMP-9, CTSK, p-AKT and AKT expression. RAW264.7 cells transfected with miR-19a-3p inhibitor-NC or inhibitor were stimulated with RANKL (50 ng/mL) for 4 days before RNA and protein collection. (I) AKT protein level and phosphorylation level in RAW264.7 cells transfected with PTEN-vector and/or miR-19a-3p mimics. Western Blot analysis was conducted after transfection 48 hours. Values are mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01.