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. Author manuscript; available in PMC: 2017 Jan 16.
Published in final edited form as: ACS Nano. 2015 Jun 2;9(7):6683–6695. doi: 10.1021/acsnano.5b00690

Figure 2.

Figure 2

RA-loaded nanovehicle induces efficient neural stem cell differentiation at low concentration. (A) Confocal laser scanning microscopy imaging of NE-4C NSC differentiation by free RA and RA-loaded nanovehicle at 1 µM (two upper panels) or 100 nM (two lower panels) concentrations. Scale bar = 50 µm. NeuN: neuron specific marker stained by Cy3-conjugated secondary antibody. (B–E) PCR analysis of effect of free RA and RA-loaded nanovehicle on inducing NE-4C NSC differentiation. The pro-neuron markers mash1 and ngn2 and mature neuron marker math2 were measured. A 100 nM RA-loaded nanovehicle was able to efficiently induce NE-4C differentiation similar to the level of 1 µM free RA. GAPDH was used as the housekeeping gene. (F) Immunofluorescence staining of βIII tubulin expression in differentiating NE-4C NSCs stimulated by 100 nM RA-loaded nanovehicle. The βIII tubulin staining was illustrated by Alexa 488-conjugated secondary antibody. Scale bar = 20 µm. (G–J) Western blot (G and H) and PCR (I and J) analysis of βIII tubulin in differentiating NE-4C NSCs stimulated by 100 nM RA-loaded nanovehicle.