Research Paper Volume 8, Issue 5 pp 899—916

Resveratrol supplementation confers neuroprotection in cortical brain tissue of nonhuman primates fed a high-fat/sucrose diet

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Figure 4. Effects of resveratrol on neuroinflammation in the cerebral cortex of HFS-fed rhesus monkeys. (A) Representative images of Iba1-positive cells in cerebral cortical brain sections (original magnification: 40x), and their respective quantification. (B) IL-6 levels in CSF. (C-K) Solubilized cerebral cortex extracts from SD-, HFS- and HFS+R-fed animals were resolved by SDS-PAGE under reducing conditions, electrotransferred onto nitrocellulose membranes and subjected to immunoblotting. All 24 brain samples were ran on a single gel (Fig. S1). (C) Representative signals associated with phosphorylated and total forms of p65Rel and that of GAPDH, which was used as loading control. (D) The ratios of phosphorylated/total p65Rel proteins are represented as scatter plots. (E) Enrichment of a select group of transcripts implicated in NF-κB signaling both in HFS vs. SD and HFS+R vs. HFS pairwise comparisons. (F) Representative signals associated with BCL2 and cleaved caspase 3, and that of GAPDH, which was used as loading control. (G) Signals associated with BCL2 and cleaved caspase 3 proteins were normalized and represented as box plots. (H) Representative immunoblot analysis for SIRT2, acetylated tubulin, and total tubulin levels. (I) Signals associated with bands of interest were normalized to Ponceau S staining and represented as box plots. (J) Representative signals associated with ALDH2 and GAPDH, the latter being used as loading control. Membrane staining was also carried out with Ponceau S dye for protein detection. The migration of molecular-mass markers (values in kilodaltons) is shown on the left of the stained membrane. (K) Signal associated with ALDH2 was normalized to GAPDH and represented as box plots. (L) Scatterplot exploring the significant association between ALDH2 and BCL2 protein expression with HFS+R feeding. SD (n=4); HFS (n=10); HFS+R (n=10). *, p < 0.05.