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Fig. 3 | Molecular Brain

Fig. 3

From: Age-related increase in caveolin-1 expression facilitates cell-to-cell transmission of α-synuclein in neurons

Fig. 3

Phosphorylation of cav-1 at tyrosine 14 is important to regulate the uptake of α-syn. a WT cav-1 OE SH-SY5Y cells were cocultured with differentiated α-syn OE SH-SY5Y cells cultured on the insert for 24 h. The samples were fixed and immunostained with pCav-1 antibody and observed under a confocal microscope. The intensity of three independent experiments was analyzed. b WT cav-1 OE SH-SY5Y cells were incubated in the presence of 1 μM α-syn monomers or α-syn fibrils (n = 5) for 10 min. The cells were then lysed, and Western blot was performed with the indicated antibodies. The band intensity of four independent experiments was analyzed. c Each stable cell line expressing EGFP only (Con), WT cav-1-EGFP (WT) and Y14A cav-1-EGFP (Y14A) was cocultured with differentiated α-syn OE SH-SY5Y cells for 5 days using a dual-chamber system (n = 5). After 24 h of coculture, the samples were stained with α-syn antibody (BD). d Each stable cell line was cocultured with differentiated A53T α-syn-mCherry OE SH-SY5Y cells (A53T-M) using the dual-chamber system (n = 4). After 24 h of coculture, the samples were stained with mCherry antibody. e Rat primary cortical neurons were transfected with EGFP, WT cav-1-EGFP and Y14A cav-1-EGFP plasmids. After one day of transfection, neurons were cocultured with A53T-M OE SH-SY5Y cells for 24 h (n = 3). The cells were stained with mCherry antibody. All images are observed under a confocal microscope. Blue indicates DAPI staining. Scale bars indicate 20 μm. The intensity of independent experiments was analyzed. *p < 0.05, **p < 0.01, ***p < 0.001, unpaired t-test (a), one-way ANOVA (be)

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