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. 2010 Feb;13(1):65–74. doi: 10.1089/rej.2009.0892

FIG. 4.

FIG. 4.

Age-related increase in proinflammatory mediators is attenuated by a lifelong calorie-restricted (CR) diet. (A) Total sciatic nerve lysates (20 μg/lane) from the indicated ages and diet were analyzed with antibodies against tumor necrosis factor-α (TNF-α, p50 and p65 subunits of NF-κB, and phospho-IκB. Tubulin is shown as a loading control. Molecular mass is shown on the left in kDa. (B) Quantification of TNF-α band intensities after normalization to tubulin is represented (##p < 0.01, Fisher protected least significant difference (PLSD) analysis; *p < 0.05, unpaired t-test, mean ± standard error of the mean [SEM], n = 3). (C) Quantification of the p50 subunit of NF-κB band intensities normalized to tubulin is shown (##p < 0.01, ###p < 0.001, Fisher PLSD analysis; *p < 0.05, unpaired t-test, mean ± SEM, n = 3). (D) Quantification of the p65 subunit of NF-κB band intensities normalized to tubulin is shown (p = 0.0546, ##p < 0.01, ###p < 0.001, Fisher PLSD analysis; *p < 0.05, unpaired t-test, mean ± SEM, n = 3). (E) Phospho-IκB levels were also analyzed within the same lysates and quantification of band intensities normalized to tubulin was performed (p = 0.0830, ##p < 0.01, ###p < 0.001, Fisher PLSD analysis; **p < 0.01, unpaired t-test, mean ± SEM, n = 3). AL, Ad libitum.