Comparative Analysis of AGE and RAGE Levels in Human Somatic and Embryonic Stem Cells under H₂O₂-Induced Noncytotoxic Oxidative Stress Conditions

Figure 4

Levels of CML, RAGE, and 20S proteasome activity in protein extracts of hES and somatic cells upon differentiation. (a) Levels of CML in protein extracts of hES (HUES3 and HUES7) and differentiated somatic cells (neural precursors, cardiac embryoids, neurons, cardiomyocytes, Hs27 fibroblast, and HUVEC) upon differentiation. Amount of protein-bound CML related to the protein load (Ponceau S staining) normalized with the Hs27 cell line. Mean ± SD, 3 separate replicates analysed per group. Statistically significant differences between groups are indicated as follows: , , and as established by one-way ANOVA and Tukey’s post hoc test. (b) Levels of RAGE relative to β-actin in protein extracts of hES and somatic cells upon differentiation, normalized with the Hs27 cell line. The quantification was performed by western blot analysis: mean ± SD, 3 separate replicates analysed per group. Statistically significant differences between groups are indicated as follows: , , , and as established by one-way ANOVA and Tukey’s post hoc test. (c) Proteasome activity: relative fluorescence units measured at 380/460 nm of total sample and inhibited fraction of AMC signal with Lactacystin 20S proteasome inhibitor, mean ± SD, 3 separate replicates analysed per group. Statistically significant differences between groups are indicated as follows: , , and as established by one-way ANOVA and Tukey’s post hoc test.

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