Fig 1: Spinal cord staining with GFAP.A: Representative photomicrographs (100x objective) of the ipsilateral ventral grey horn of L3-L6 spinal cords sections from all groups immunostained for GFAP (GFAP (GA-5): sc-58766, Mouse Monoclonal–Santa Cruz Biotechnology) at week 3 (A1, A3, A5, A7 and A9) and week 6 (A2, A4, A6, A8 and A10) following nerve crush injury. The sham group (n = 12) (A1 and A2) show normal and clear GFAP-immunoreactive astrocytes in the ventral horn. Whereas, the crush group (n = 12) (A3 and A4) display a remarkable increase in GFAP immunoreactive astrocytes. The TEGBE (n = 12) (A5, A6), G-B (n = 12) (A7, A8) and GBE-treated (n = 6) (A9, A10) groups show an observable decrease in the GFAP-immunoreactive astrocytes compared to crush at the week 3 and week 6 post-injury. Black arrows indicate the GFAP immunoreactive astrocytes. Scale bar = 200 µm. B, C: Bar graphs showing the average number of total GFAP immunoreactive neurons in the ventral grey horn (B) dorsal grey horn (C) and obtained from the different experimental groups at week 3 and week 6 following sciatic nerve injury. Data show a significant decrease in the number of the GFAP-immunoreactive astrocytes in the ventral and dorsal grey matters at week 3 and week 6 post-injury. B: Note the significant increase in the GFAP immunoreactive astrocytes in the ventral horn in crush compared to sham and treated groups at week 3 and week 6 post-injury. * indicates p<0.0001, crush group vs. sham group, TEGBE and G-B treated groups. ** indicates p<0.0001, GBE treated group vs. sham group and G-B-treated group. ? indicates p<0.002, TEGBE-treated group vs. sham group and G-B-treated group. O indicates p<0.0001, crush vs. sham group. ? indicates p<0.005, crush vs. all treated groups. C: Note the significant increase in GFAP immunoreactive astrocytes in the dorsal horn in crush compared to sham, TEGBE and G-B and GBE-treated groups at week 3 and week 6 post-surgery. * indicates p<0.0001, crush vs. sham group, TEGBE and G-B-treated groups. ? indicates p<0.02, crush compared to GBE treated group. ? indicates p<0.0001, GBE treated group vs. sham group and G-B-treated group. ? indicates p<0.02, GBE-treated group vs. TEGBE-treated group. ** indicates p<0.009, GBE-treated group vs. sham group. O indicates p<0.05, TEGBE-treated group vs. sham group. Data represent mean ± SD (n = 6 /group). D: Illustrates the Western blot graph and the statistical data analysis of the GFAP expression in the spinal cords obtained from all the experimental groups except GBE treated group due to a shortage of animal supply. In each group, the lumbar spinal cords from three rats were analyzed at week 3 and week 6 post-injury. The antibody used to detect the GFAP (Anti-GFAP antibody (ab7260)–Abcam Biochemical®, Cambridge, MA, USA). The bar graph shows the GFAP density in spinal cord samples (n = 3 rats/group) normalized with densities of actin band (Molecular weight-42). The labeled GFAP bands were quantified. Note that the crush group exhibits a significant (*p<0.0001) increase in the GFAP expression compared to sham and naive (n = 6) groups. Whereas the TEGBE and G-B treated groups show a substantial decrease in GFAP immunoblotting, but still significantly (**p<0.001) higher compared to naive and sham animals at weeks 3 and 6 post-injury.
Fig 2: Analysis of GFAP expression and TUNEL in brain tissue sections from Eif5a or Dhps CKO mice.A, the GFAP IHC was performed using the anti-GFAP primary antibody (Abcam, ab7260, 1:1000 dilution) as described under Experimental procedures with the brain slides of Eif5aCamk2a, DhpsEmx1, and DhpsCamk2a mice and their respective controls, at different time points after birth. B, TUNEL assays were performed as described in Experimental procedures with the brain slides of Eif5aEmx1, DhpsEmx1, Eif5aCamk2a, and DhpsCamk2a mice and their respective controls, at different time points after birth. CKO, conditional KO; GFAP, glial fibrillary acidic protein; IHC, immunohistochemistry; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.
Supplier Page from Abcam for Anti-GFAP antibody