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. 2016 Apr 19;7(21):29989–30002. doi: 10.18632/oncotarget.8802

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Copyright: © 2016 Moon et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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A. ZW 13-2 and Zpl 3-4 cells were incubated with PrP (106-126) in a dose-dependent manner for 24h. Cell viability was measured by annexin V assay. Cells were treated with FITC-annexin V and PI, which binds to phosphatidylserine to the plasma membrane and nuclei during apoptosis. B. Bar graph indicating the average number of annexin V negative cells. C. Representative immunofluorescence images of TUNEL-positive (green) ZW or Zpl cells at 24 h after exposure to 100 μM of PrP (106-126). The cells were counterstained with PI (red) to show all cell nuclei. D. Lactate dehydrogenase (LDH) assay was used to quantify LDH released into the medium. E. ZW 13-2 and Zpl 3-4 cells were incubated with PrP (106-126) in a dose-dependent manner for 6h. The incubated cells were assessed for LC3B production, prion protein and P62 expression by western blot analysis. β-actin was used as loading control. Bar graph indicating the average values of p62 expression levels F.. G. ZW 13-2 and Zpl 3-4 cells were stained with rabbit anti-p62 (red) and DAPI (nuclei, blue) for immunocytochemistry using confocal microscopy. *** p < 0.001; significant differences between each treatment group. PrP, Prion peptide (106-126); PrPc, normal prion protein; adj.volume, adjustment of volume (band volume minus background volume).

A. ZW 13-2 and Zpl 3-4 cells were incubated with PrP (106-126) in a dose-dependent manner for 24h. Cell viability was measured by annexin V assay. Cells were treated with FITC-annexin V and PI, which binds to phosphatidylserine to the plasma membrane and nuclei during apoptosis. B. Bar graph indicating the average number of annexin V negative cells. C. Representative immunofluorescence images of TUNEL-positive (green) ZW or Zpl cells at 24 h after exposure to 100 μM of PrP (106-126). The cells were counterstained with PI (red) to show all cell nuclei. D. Lactate dehydrogenase (LDH) assay was used to quantify LDH released into the medium. E. ZW 13-2 and Zpl 3-4 cells were incubated with PrP (106-126) in a dose-dependent manner for 6h. The incubated cells were assessed for LC3B production, prion protein and P62 expression by western blot analysis. β-actin was used as loading control. Bar graph indicating the average values of p62 expression levels F.. G. ZW 13-2 and Zpl 3-4 cells were stained with rabbit anti-p62 (red) and DAPI (nuclei, blue) for immunocytochemistry using confocal microscopy. *** p < 0.001; significant differences between each treatment group. PrP, Prion peptide (106-126); PrPc, normal prion protein; adj.volume, adjustment of volume (band volume minus background volume).