Figure 3. Inhibition of endogenous levels of GLS2 reduce neurite outgrowth induced by RA. (A) Representative images of neurite outgrowth in the SHSY-5Y cell line. Cells were transfected with scramble + GFP or siGLS2 + GFP and untreated or treated with retinoic acid (RA). (B) After 48 h cells were fixed and analyzed for neurite extension as described in Methods. Data represent mean ± s.d. of 3 different experiments. Glutamine withdrawal impairs in vitro terminal differentiation of cortical neurons. (C) Cortical neurons were cultured either in the presence or absence of glutamine and fixed and stained for the indicated proteins after 7 days. Magnification 40×. (D) Quantification of fluorescence intensity of the neuronal marker β-III-Tubulin and of the indicated synaptic proteins was performed using Zeiss LSM 510 software analysis.