Extended Data Fig. 5: Aβ40 induces cellular senescence of RAW264.7 and human macrophages.
(a-b) RAW264.7 cells were subjected to in vitro experiments and treated with Aβ40 (20 μg/ml) for the indicated time period. (a) Telomere length was assessed using a telomere probe (C-rich) (TELC) and flow cytometry. Data are presented as mean values +/- SEM. Experiments were repeated four times. **P < 0.001, ***P < 0.001, by one-way ANOVA, compared with the 0h group. (b) RAW264.7 treated with Aβ40 (20 μg/ml) for indicated time period were subjected to immunostaining for LMNB1, HMGB1, PLIN2. Experiments were conducted three times. Representative images were displayed. Single channel images were shown in Supplementary Fig. 4b. (c-f) Human macrophages were derived from blood monocytes donated by healthy young adults (age = 20–30 years) using human serum and MCSF. (c) Human macrophages were treated with PBS for indicated time points then subjected to analysis of LMNB1, HMGB1, p-γH2AX, pSTAT3, and SA-β-Gal, followed by co-staining with phalloidin. Single channel images were shown in Supplementary Fig. 4c. Experiments were conducted four times. (d) The cell cycle of the human macrophage cultures was analyzed using Ki67 and DAPI staining via flow cytometry. (e) Telomere length was assessed using a telomere probe (C-rich) (TELC) and flow cytometry. (f) The cultures were labeled with WGA-AF488, and migrasomes produced by macrophages were quantified.
