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. 2020 Aug 31;8:712. doi: 10.3389/fcell.2020.00712

FIGURE 2.

FIGURE 2

Citicoline reduces the expression of apoptotic factors in cochlear HCs after neomycin exposure. (A) Immunofluorescence staining with TUNEL and Myo7A in the middle turn of the cochlea after different treatments. (B) Quantification of the numbers of TUNEL and Myo7A double-positive cells in (A). (C) Immunofluorescence staining of cleaved caspase 3 and Myo7A in the middle turn of the cochlea after different treatments. (D) Quantification of the numbers of cleaved caspase 3 and Myo7A double-positive cells in (C). (E) Western blot showing that the amount of cleaved caspase 3 in the neomycin-only group was higher than in the undamaged control. The amount of cleaved caspase 3 induced by neomycin was significantly reduced by pretreatment with citicoline. (F) Quantification of the western blot in (E). (G) The mRNA levels of five apoptosis-related genes were analyzed by qRT-PCR after neomycin and citicoline treatment normalized to GAPDH and presented as the fold change compared to control levels. Data are shown as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars = 20 μm.