Figure 2
Spermine treatment clears prion aggregates in SMB and CAD cells terminally differentiated into neurons and reduces ROS in CAD cells. SMB.s15 and CAD22L cells were treated with 5 μM spermine for the mentioned period. (a) Representative immunofluorescence images showing reduced PrPSc staining after 24 and 72 hrs of spermine treatment in SMB.s15 cells (N = 3). (b) Significantly reduced pixel intensity of PrPSc in immunofluorescence images on 72 hrs spermine treatment in SMB.s15 cells. (c) Proteinase-K treated lysates immunoblotted for PrPSc after 0, 24 and 72 hrs of treatment with spermine in SMB.s15 cells (N = 3). (d) Significantly reduced relative density of total PrPSc after 72 hrs of spermine treatment. (e) Significantly reduced relative density of di- and mono- glycosylated forms of PrPSc after 72 hrs of spermine treatment. (f) CAD22L cells showing reduced immunofluorescence for PrPSc after 72 hrs of spermine treatment (N = 2). (g–i) Reduced expression of total PrPSc and PrPSc glycoforms in CAD22L cells, after 72 hrs of spermine treatment (N = 2). PrPSc glycoforms are shown with red arrows. (j) Reduced DCF fluorescence in CAD22L cells after 72 hrs of spermine treatment (N = 2). (k) Significantly reduced DCF mean intensity in CAD22L cells. Scale bar = 10 μm. Respective full length blots are shown in Supplementary Fig. 3. Abbreviations used: Chloroquine (CQ), Transmission Electron Microscopy (TEM), Dichlorofluorescein (DCF).
