Fig. 9: 4-PBA rescues protein synthesis and p-eIF2α in 3Tg i-Astro and tubulogenesis in pericyte/EC co-cultures. | Cell Death & Disease

Fig. 9: 4-PBA rescues protein synthesis and p-eIF2α in 3Tg i-Astro and tubulogenesis in pericyte/EC co-cultures.

From: Protein synthesis inhibition and loss of homeostatic functions in astrocytes from an Alzheimer’s disease mouse model: a role for ER-mitochondria interaction

Fig. 9

a WT- and Tg-iAstro were treated or not with 4-PBA 3 µM, for 48 h, cells were pulsed with puromycin 4 µM and analysed by WB with anti-puromycin antibody and ponceau staining. Data are expressed as mean ± SEM from 6–4 independent experiments; **p < 0.01; ***p < 0.001, one-way ANOVA, Sidak’s multiple comparisons. b WB analysis of eIF2α phosphorylation on WT-iAstro and 3Tg-iAstro, treated or not with 4-PBA 3 µM, for 48 h. Data are expressed as mean ± SEM from four independent experiments; **p < 0.01, one-way ANOVA, Sidak’s multiple comparisons. c Representative images and quantification of SPLICS fluorescence, indicating ER-mitochondrial contacts at ∼8–10 nm distance, in WT-iAstro, 3Tg-iAstro, and in 3Tg-iAstro treated with 4-PBA (3 µM, for 48 h). Data are expressed as mean ± SEM of n = 22 (WT-iAstro), n = 30 (Tg-iAstro), n = 32 (Tg-iAstro + 4-PBA), from three independent coverslip, ****p < 0.0001, one-way ANOVA, Sidak’s multiple comparisons. Scale bar = 20 μM. d Co-cultures of pericytes, endothelial cells and WT-iAstro or 3Tg-iAstro (pre-treated or not with 4-PBA for 48 h) were plated in a layer of Matrigel in presence or absence of 4-PBA (3 µM). After 8 h, bright field images were taken using a Zeiss 710 confocal laser scanning microscope, scale bar = 500 μm. Data are expressed as mean ± SEM, n = 4 from two independent experiments; ***p < 0.001 by one-way ANOVA, Sidak’s multiple comparisons.

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