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. 2014 Apr 16;9(4):e94035. doi: 10.1371/journal.pone.0094035

Figure 7. Dynamic localization of GKRP in cultured tanycytes in response to glucose.

Figure 7

A–D. GKRP immunolocalization (red) in tanycytes preincubated with 0.5 mM glucose for 6 h (A–B) or incubated with 15 mM glucose for 30 min (C–D). Nuclei (A–C) and Vimentin (B, D) are stained in blue and green, respectively. Arrows and arrowheads show nuclear and cytoplasmic GKRP localization, respectively. Images were obtained by Z-stack reconstruction, and white lines indicate orthogonal planes (XZ and YZ) shown in the lateral and lower panels. E, Nuclear intensity of GKRP immunofluorescence in tanycytes cultured in the presence of 0.5 mM (lane 1) and 15 mM (lane 2) glucose. The results represent the mean±sd of 200 cells from three independent primary cultures. F, Immunoblots of GKRP (69 kDa, upper panel) and the nuclear marker, lamin-B1 (68 kDa, lower panel), in nuclear extracts obtained from cells cultured with 0.5 mM (lane 1) and 15 mM (lane 2) glucose. G, GKRP nuclear expression levels assessed by western-blot were quantified and normalized with lamin-B for cells cultured with 0.5 mM (lane 1) and 15 mM (lane 2) glucose. The data shows the means±SD from six independent experiments. * p<0.05; ** p<0.01. Scale bar in A-D, 50 µm.