Figure 3: Effect of rosuvastatin and betulin on neuroserpin aggregation and cholesterol level.
From: Cholesterol impairment contributes to neuroserpin aggregation
(a) Hela-GFP and Hela cells were mixed 1:1, treated with 0.1 μM rosuvastatin or 20 μM betulin for 8days, fixed in 3.7% paraformaldeide and incubated with the primary antibody anti-neuroserpin (Ab32901, Abcam) coniugated PLA probes (1:50), overnight at 4 °C. The presence of aggregates are visualized using Duolink in Situ staining according to the manufacturer’s instructions. Nuclei are stained with DAPI. Images are acquired by Leica SP2 laser scanning confocal microscope. Each red spot represents a neuroserpin aggregate. The number of spots per cell for each condition in the images showed in this panel is quantified as described in the methods section. (b) Untreated or treated cells with 0.1 μM rosuvastatin or 20 μM betulin (8 days) were fixed with 3.7% paraformaldeide and then incubated with TNM-AMCA (1 μM) to stain cholesterol for 1 h at room temperature. Images were acquired by Leica SP2 laser scanning confocal microscope.
