rabbit polyclonal anti phospho ampk t172  (Cell Signaling Technology Inc)

Journal: Molecules and Cells

Article Title: CpG oligodeoxynucleotide reduces PrP Sc accumulation and prolongs survival in prion-infected mice

doi: 10.1016/j.mocell.2026.100335

Figure Lengend Snippet: CpG ODN induces AMPK signaling and attenuates phosphorylated p62 accumulation in 22L scrapie-infected mice. (a) Western blot analysis of brain lysates at 170 dpi showing expression levels of p-AMPK T172, AMPK, p-ULK1 S555, ULK1, p-p62 S403, p62, ATG12–5, and LC3 I/II in brains of 22L scrapie-infected mice with or without CpG ODN at 170 dpi. (b) Relative intensity of p-AMPK, p-ULK, p62, p-p62, ATG12–5, and LC3-II represented as bar graphs (mean ± S.E.M, n = 6). Statistical significance was determined by 1-way ANOVA with Tukey’s post hoc test. * P < .05, ** P < .01, *** P < .001. NS: not significant.

Article Snippet: Membranes were blocked with 5% nonfat dry milk in PBST (8 mM Na 2 HPO 4 , 2 mM KH 2 PO4, 138 mM NaCl, 2.7 mM KCl, 0.1% Tween 20; pH 7.4) for 1 hour at room temperature (RT), followed by overnight incubation at 4 °C with the following primary antibodies: mouse monoclonal anti-PrP 3F10 (1:2000) , rabbit polyclonal anti-TLR9 (1:2000, Abcam, Cambridge, UK), rabbit polyclonal anti-phospho AMPK T172 (p-AMPK T172) (1:2000, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal anti-AMPK (1:2000, Cell Signaling Technology), rabbit polyclonal anti-phospho-p62 S403 (p-p62 S403) (1:2000, Cell Signaling Technology), rabbit monoclonal anti-p62 (1:2000, MBL, Nagoya, Japan), rabbit monoclonal anti-phospho-ULK1 S555 (p-ULK1 S555)(1:2000, Cell Signaling Technology), rabbit polyclonal anti-ULK1 (1:2000, Cell Signaling Technology), rabbit polyclonal anti-LC3 I/II (1:2000, Cell Signaling Technology), rabbit polyclonal anti-GFAP (1:2000, CosmoBio, Tokyo, Japan), rabbit polyclonal anti-ATG12 (1:2000, Cell Signaling Technology) and mouse monoclonal anti-β-actin (1:2000, Sigma-Aldrich).

Techniques: Infection, Western Blot, Expressing

Journal: Molecules and Cells

Article Title: CpG oligodeoxynucleotide reduces PrP Sc accumulation and prolongs survival in prion-infected mice

doi: 10.1016/j.mocell.2026.100335

Figure Lengend Snippet: CpG ODN reduces PrP Sc accumulation and activates AMPK-associated signaling in 22L scrapie-infected neuronal cells. (a and b) Immunoblot analysis of (a) PrP Sc and total PrP, and (b) TLR9, p-AMPK T172, total AMPK, p-ULK1 S555, total ULK1, ATG12–5, total p62, and LC3 I/II in 22L scrapie-infected neuronal cells (ZW-22L) treated with CpG ODN (0, 1, or 3 µM) for 6 hours. For PrP Sc detection, equal amounts of protein were incubated with proteinase K (2 µg/ml) for 1 hour. (c) Densitometric quantification of p-AMPK, p-ULK1, ATG12–5, total p62, and LC3-II levels. Data are presented as mean ± S.E.M ( n = 3). (d) Immunoblot analysis of PrP Sc , p-AMPK T172, and total AMPK in ZW-22L cells treated with CpG ODN (3 µM, 6 hours) in presence or absence of the TLR9 antagonist ODN 2088 (5 µM, 7 hours). Data represents 3 independent experiments ( n = 3). Statistical significance was determined by 1-way ANOVA with Tukey’s post hoc test (* P < .05, *** P < .001).

Article Snippet: Membranes were blocked with 5% nonfat dry milk in PBST (8 mM Na 2 HPO 4 , 2 mM KH 2 PO4, 138 mM NaCl, 2.7 mM KCl, 0.1% Tween 20; pH 7.4) for 1 hour at room temperature (RT), followed by overnight incubation at 4 °C with the following primary antibodies: mouse monoclonal anti-PrP 3F10 (1:2000) , rabbit polyclonal anti-TLR9 (1:2000, Abcam, Cambridge, UK), rabbit polyclonal anti-phospho AMPK T172 (p-AMPK T172) (1:2000, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal anti-AMPK (1:2000, Cell Signaling Technology), rabbit polyclonal anti-phospho-p62 S403 (p-p62 S403) (1:2000, Cell Signaling Technology), rabbit monoclonal anti-p62 (1:2000, MBL, Nagoya, Japan), rabbit monoclonal anti-phospho-ULK1 S555 (p-ULK1 S555)(1:2000, Cell Signaling Technology), rabbit polyclonal anti-ULK1 (1:2000, Cell Signaling Technology), rabbit polyclonal anti-LC3 I/II (1:2000, Cell Signaling Technology), rabbit polyclonal anti-GFAP (1:2000, CosmoBio, Tokyo, Japan), rabbit polyclonal anti-ATG12 (1:2000, Cell Signaling Technology) and mouse monoclonal anti-β-actin (1:2000, Sigma-Aldrich).

Techniques: Infection, Western Blot, Incubation