Figure 3
The infection of sh-Src in the ARC area reduced Src expression in this area. Images (left and right) in (a) recorded from the same field of cultured ARC neurons show lentivirus-mediated GFP and sh-Src infection; Bar: 100 µm. Arrowheads indicate infected neurons. A standard diagram in (b) (left) indicates the coronal section at 3.3 mm posterior to Bregma39. A fluorescence image in (b) (right) was recorded from the ARC area indicated within the red square on the diagram (left). The NeuN antibody co-labeling of ARC neurons is shown in Fig. S1. Gels shown in (c) and (d) were respectively loaded with lysates prepared from the ARC area of naïve animals, and animals at days 3 and 7 after the infusion of sh-Src (c) or sh-NC (d) into the ARC area. Gels shown in (e) were respectively loaded with lysates prepared from the ARC area of naïve animals and animals at day 7 after intra-ARC infusion of sh-Src. Gels shown in (f) were respectively loaded with lysates prepared from the cerebral cortex (left blots) and the dorsal horn of the lumbar spinal cord (SDH, right blots) of naïve animals and animals at day 7 after the intra-ARC infusion of sh-Src. Each group of blots was cropped from a same filter, and probed with antibodies as indicated on the left side of blots. An example demonstrating the probe of a full length filter is shown in Fig. S6. The ratio of the band intensity of Src, Fyn or Lyn versus that of actin or GAPDH was calculated. Bar graphs in (c–f) respectively show the summary data (mean ± SEM) of ratios normalized to those in naïve animals (=1, dashed line). *p < 0.05, Dunnett’s post hoc test in one-way ANOVA in comparison with that of naïve rats. Values in brackets indicate the number of experimental repeats.
