Skip to main content
. 2019 Sep 20;13:415. doi: 10.3389/fncel.2019.00415

FIGURE 3.

FIGURE 3

The suppressing effects of ivabradine (IVA) on the mRNA and protein expressions of HCN channel isoforms and GAP-43 and the neurite outgrowth. (A–D) Real-time PCR and western blotting results of the mRNA and protein expression levels respectively for HCN1-4 isoforms in NGF-treated PC12 cells relative to GADPH level. ∗∗p < 0.01, ∗∗∗p < 0.001 vs. NGF treatment alone in the mRNA level. #p < 0.05, ##p < 0.01 vs. NGF treatment alone in the protein level. (E) Representative western blotting bands (upper) and the mRNA and protein semiquantitative values (lower) of GAP-43 in NGF-treated PC12 cells. ∗∗p < 0.01, ∗∗∗p < 0.001 vs. NGF treatment alone in the mRNA level. #p < 0.05, ###p < 0.001 vs. NGF treatment alone in the protein level. (F) Immunofluorescent stains of GAP-43 protein (red), displaying that NGF induced, while ivabradine inhibited, the neurite outgrowth in differentiating PC12 cells. DAPI was used for cell nuclei staining (blue). Scale bar, 10 μm. (G,H) Quantitative morphological parameters of neurite outgrowth including the total neurite length and the maximal neurite length (longest process per cell). Results are calculated from three independent experiments and presented as mean ± SEM, 96 < n < 213, ∗p < 0.05, ∗∗p < 0.01 vs. NGF treatment alone.