Fig. 1
A-E DI TNC1 cells were treated with 1 µM Aβ peptide for 24 h and compared to untreated and positive controls. A Western blotting (WB) reveals an increase of GFAP on protein level after exposure to Aβ (t-test, p=0.0183, n=3). B, C Immunocytochemistry (ICC) labeling of GFAP after exposure to Aβ confirms an increase seen by WB (t-test, p<0.0001, n=15) (AFU: Absolute Fluorescence Units). C Exemplary images of GFAP signals after treatment. D, E CellRox® immunoreactivity significantly increases after Aβ treatment, indicating higher ROS levels. H2O2 was used as a positive control to confirm the sensitivity of the fluorescent marker. As expected, adding H2O2 also significantly increases CellRox® fluorescence intensity (one-way ANOVA, p=0.0068; Tukey posthoc test: Ctrl vs. H2O2 p=0.0067; Ctrl vs. Aβ p=0.044, n=10). E Exemplary images of CellRox® fluorescence after treatment. F-H Detection of PTFAR in astrocytes on mRNA level using qRT-PCR (F) and protein levels using ICC (G) and WB (H). F ptafr mRNA was detected at a level of 1.62% of the housekeeping gene hmbs in DI TNC1 astrocytes. G Anti-PTAFR antibodies show positive labeling in DI TNC1 astrocytes. H The same anti-PTAFR antibodies used in ICC (G) reveal a protein band at the expected size for PTAFR in WB. C, E, G Scale bar = 30 μm