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. 2020 Jun 24;25(12):2907. doi: 10.3390/molecules25122907

Figure 6.

Figure 6

AN07 attenuates methylglyoxal (MG)-induced neurite damage in SH-SY5Y cells. (A,B) Representative images showed the protective effect of AN07 on MG-induced reduction in neurite outgrowth in SH-SY5Y cells. Cells were incubated with retinoic acid (10 μM) for five days to induce differentiation. Differentiated SH-SY5Y cells were pre-treated with AN07 (0.01–1.0 μM) for 1 h and then exposed to 600 μM MG for another 24 h. Neurites were imaged with a phase-contrast microscope (A), and the neurite length of cells (B) was calculated using the ImageJ software. Scale bar, 100 μm. Columns, mean ± S.E.M. from at least 30 cells of three independent experiments. (C,D) AN07 down-regulates Rho-associated protein kinase 2 (ROCK2) expression and LIM kinase 1 (LIMK1) phosphorylation in MG-treated SH-SY5Y cells. Upper panels, representative immunoblots. Lower panels, densitometry analyses of the relative ratio of ROCK2/glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein and phosphorylated-LIMK1/total-LIMK1 (t-LIMK1), which is represented as percentages of the control group. Columns, mean ± S.E.M. from at least three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.