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. 2014 Oct 14;21(1):61–70. doi: 10.1111/cns.12334

Figure 2.

Figure 2

Potentiation of NGF‐induced neuritogenesis by puerarin. (A) Representative images of PC12 cells with or without neurites. PC12 cells were treated with puerarin alone or in combination with NGF (2 ng/mL). The cells were stained by neurite outgrowth staining kit and examined under a Zeiss fluorescence microscope. Scale bar, 100 μm. (B) Puerarin potentiates NGF‐induced neuritogenesis in a concentration‐dependent manner. The neurite‐bearing cells were enumerated with NIH Image J. Untreated PC12 cells were used as control. The values represent the means ± SD (n = 3). **P < 0.01; *** P < 0.001 (sample vs NGF 2 ng/mL). (C) Immunofluorescence staining of neuronal biomarker MAP2. The cells were treated with puerarin and NGF for 72 h as follows: A, control; B, NGF (2 ng/mL); C, puerarin (50 μM); D, NGF (2 ng/mL) and puerarin (50 μM). Following staining with anti‐MAP2 and DAPI, the cells were examined under a Zeiss fluorescence microscope. (D) Western blot analysis of neuronal biomarker β3‐tubulin in PC12 cells. The cells were treated with vehicle control, NGF (2 ng/mL), puerarin (10 μM) or combination of NGF (2 ng/mL) and puerarin (10 μM) for 72 h. β3‐tubulin expression was analyzed by Western blotting using specific antibody, whereas GAPDH was analyzed as the control of protein loading.