Fig. 4
A-E DI TNC1 cells were treated with 1 µM Aβ peptide for 24 h, and Aβ with SPL and YPL, and compared to untreated controls and positive controls. A Western blotting (WB) reveals an increase of GFAP on protein levels normalized to β-ACTIN (Ctrl, Aβ, and Aβ+SPL) or VINCULIN (Ctrl, and Aβ+YPL) after exposure to Aβ. The results are shown as % of respective Ctrl. The increase is normalized by SPL and YPL treatment (one-way ANOVA, p<0.0001; Tukey post-hoc test: Ctrl vs. Aβ p=0.0173; Aβ vs. Aβ+SPL p=0.0046; Aβ vs. Aβ+YPL p<0.001; Ctrl vs. Aβ+YPL p=0.0004; Aβ+SPL vs. Aβ+YPL p=0.0038, n=3). B, C Immunocytochemistry (ICC) labeling of GFAP after exposure to Aβ confirms the effects of SPL and YPL. The increase in GFAP induced by Aβ is significantly reduced in SPL and YPL-treated astrocytes (one-way ANOVA, p<0.0001; Tukey posthoc test: Ctrl vs. Aβ p<0.0001; Aβ vs. Aβ+SPL p<0.0001; Aβ vs. Aβ+YPL p<0.001; Ctrl vs. Aβ+SPL p<0.0001; Ctrl vs. Aβ+YPL p<0.0001, n=15). (AFU: Absolute Fluorescence Units). C Exemplary images of GFAP signals after treatments. D, E CellRox® immunoreactivity significantly increases after Aβ treatment, indicating higher ROS levels. The increase in ROS induced by Aβ is significantly reduced in SPL and YPL-treated astrocytes (one-way ANOVA, p=0.0068; Tukey posthoc test: Ctrl vs. H2O2 p=0.0015; Ctrl vs. Aβ p=0.0254; Aβ vs. Aβ+SPL p<0.0001; Aβ vs. Aβ+YPL p<0.001; Ctrl vs. Aβ+SPL p<0.0001; Ctrl vs. Aβ+YPL p<0.0001, n=10). E Exemplary images of CellRox® fluorescence after treatments. C, E Scale bar = 30 μm