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. 2020 May 9;12(5):1356. doi: 10.3390/nu12051356

Figure 4.

Figure 4

Glial Markers. Immunochemical analysis of the frontal cortex (A) and hippocampus (B) processed for anti-glial fibrillary acidic protein (GFAP). To investigate more than GFAP protein on the same blot, both membranes of the frontal cortex and hippocampus were stripped and incubated with anti-GAPDH antibody (loading control). Control images are re-used for illustrative purposes. The densitometric analysis of bands is expressed as the ratio between the optical density of protein and reference protein (GADPH) where the value for LZRs of different ages is set as 1. Blots are representative of one of three separate experiments. (C) Sections of the frontal cortex of LZRs and OZRs of different weeks of 12-, 16-, and 20-weeks age processed for the immunohistochemistry against the GFAP. V; IV: fifth and sixth layers of the frontal cortex. Magnification 40X. Calibration bar: 25 μm. (D) Sections of the hippocampus (CA1 subfield) of LZRs and OZRs of different weeks of age processed for the immunohistochemistry against ionized calcium-binding adaptor molecule 1 (Iba-1) to detect microglial cells O: stratum oriens; P: pyramidal neurons; R: stratum radiatum. Calibration bar: 25 μm. (E) Sections of the hippocampus (CA1 subfield) and (F) dentate gyrus of 20 weeks-old LZRs and OZRs processed for double immunohistochemistry against Iba-1 (Alexa Fluore 594) and GFAP (Alexa Fluore 488). DAPI was used to counterstain nuclei. O: stratum oriens; P: pyramidal neurons; R: stratum radiatum. Magnification 40X. Calibration bar: 25 μm.