2. MicroRNA-9 (miR-9) induces differentiation of bone marrow mesenchymal stem cells (BMSCs).
Autophagy contributes to the efficiency of neuronal differentiation in BMSCs.
Does miR-9 activate autophagy?

3. Lower percentage of cells presenting NSE and MAP-2 (miR-9-, purple)
miR-9 induced neuronal cell differentiation in BMSCs
Relative miR-9 expression
(Real-time PCR)
Higher miR-9 expression in cells transfected with MiR-9-1-LV (miR-9+; red)
Higher percentage of cells presenting neuronal cell markers NSE and MAP-2 (miR-9+, orange)
Lower miR-9 expression in cells transfected with Anti-mmu-miR-9-1 inhibitor (miR-9-; blue)
% of neuronal marker positive cells
Lower percentage of cells presenting NSE and MAP-2 (miR-9-, purple)
Western Blot assay

4. Autophagy activation by miR-9:
Immunofluorescent staining of autophagic marker LC3 in differentiated BMSCs
White arrows point at LC3 positive dots
Higher expression of miR-9 (miR-9+)
LC3 positive dots/cell
More positive dots/cell of autophagic marker LC3 (A1; red)
Lower expression of miR-9 (miR-9-)
Less positive dots/cell of autophagic marker LC3 (A3; blue)

5. Autophagy activation by miR-9: Western Blot and LC3-II/LC3-I ratio
Western blot assay
Higher ratio of LC3-II/LC3-I
Higher degree of autophagosome formation
Higher expression of miR-9 (miR-9+)
Higher LC3-II expression and LC3-II/LC3-I ratio (red)
Lower expression of miR-9 (miR-9-)
Lower LC3-II expression and LC3-II/LC3-I ratio (blue)

6. Conclusion
miR-9 can promote neuronal differentiation in BMSC cells by modulating autophagic activity
miR-9 could be used to further the study of the mechanisms of differentiation
It could also be used to promote BMSC-based therapy of diseases affecting the nervous system
However, the downstream mechanism of this autophagic modulation activity was not explored and necessitates more study