Figure 4.

Overexpression of miR-135a-5p Promotes Neurite Outgrowth in PC12 Cells

(A) Neurite length in PC12 cells transfected with 135a mimic, 135a inhibitor, siROCK, or their NC (scale bar, 100 μm; n = 100). (B) The PC12 cells transfected with the miR-135a-5p mimic group had a significant increase in neurite extension compared to those transfected with the NC group; Cells transfected with the miR-135a-5p inhibitor showed a significant decrease in neurite length (n = 100). (C and D) RNA levels of ROCK1 (C) and ROCK2 (D) after siRNA transfection in cells (n = 3). (E) Protein levels of ROCK1/2, Ser473-pAKT/AKT, Thr308-pAKT/AKT, and Ser9-pGSK3β/GSK3β (n = 3). (F) The expression levels of ROCK1 and ROCK2 were significantly decreased in miR-135a-5p mimic group, whereas the miR-135a-5p inhibitor group increased ROCK1 and ROCK2 levels; The levels of pAKT (Ser473 and Thr308) and pGSK3β (Ser9) were significantly higher in miR-135a-5p mimic group; Moreover, the miR-135a-5p inhibitor group reduced the levels of pAKT (Ser473 and Thr308) and pGSK3β (Ser9) (n = 3). (G) Protein levels of ROCK1/2, Ser473-pAKT/AKT, Thr308-pAKT/AKT, and Ser9-pGSK3β/GSK3β (n = 3). (H) the reduced levels of pAKT (Ser473 and Thr308) and pGSK3β (Ser9) induced by the miR-135a-5p inhibitor were rescued by the downregulation of ROCK (n = 3). Statistical data were measurement data, described as mean ± standard deviation. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. NC, negative control; 135a, miR-135a-5p; siROCK, siRNA targeting ROCK1 and siRNA targeting ROCK2.