Journal: PLoS ONE

Article Title: Neuroprotective Effect of TAT-14-3-3ε Fusion Protein against Cerebral Ischemia/Reperfusion Injury in Rats

doi: 10.1371/journal.pone.0093334

Figure Lengend Snippet: (A) Representative electron microphotographs showing autophagosomes in the ischemic penumbra of cerebral cortex of sham, vehicle or TAT-14-3-3ε pre-treated animals 24 h after reperfusion. Autophagosomes are indicated by arrows. Scale bar = 1 μm. (B) Western blot analysis for the expression of LC3B, p62 and Beclin-1 in cerebral hemisphere. (C, D and E) Quantitation of LC3B-II, p62 and Beclin-1 expression, respectively, normalized to β-actin. The data are presented as the mean ± SD (n = 5 per group). # p <0.05 and ## p <0.01 versus sham group. * p <0.05 versus vehicle group.

Article Snippet: Polyclonal rabbit anti-LC3B antibody (1∶1000, Santa Cruz, CA, USA), polyclonal rabbit anti-Beclin-1 antibody (1∶1000, Santa Cruz, CA, USA) and polyclonal rabbit anti-p62/SQSTSM1 antibody (1∶500, Abgent, CA, USA) were used for immunoreactivity detection.

Techniques: Western Blot, Expressing, Quantitation Assay

Journal: PLoS ONE

Article Title: Neuroprotective Effect of TAT-14-3-3ε Fusion Protein against Cerebral Ischemia/Reperfusion Injury in Rats

doi: 10.1371/journal.pone.0093334

Figure Lengend Snippet: Rats were treated with an i.c.v of saline or 3-MA (600 nmol) at the end of 2h ischemia, or RAP (35 pmol, i.c.v.) combined with TAT-14-3-3ε (10 mg/kg, i.v.) 2 h before 2 h MCAO. Rats were then subjected to 24 h reperfusion, after which all animals were sacrificed. (A) Western blot analysis of the LC3B, p62 and Beclin-1 protein expression in the ischemic cerebral hemisphere. (B, C and D) Quantitation of LC3B-II, p62 and Beclin-1 expression, respectively, normalized to the loading control (β-actin). Bar represents mean ± SD (n = 4 per group). ## p <0.01 compared with sham group. * p <0.05 compared with vehicle group. § p <0.05 compared with RAP + TAT-14-3-3ε group.

Article Snippet: Polyclonal rabbit anti-LC3B antibody (1∶1000, Santa Cruz, CA, USA), polyclonal rabbit anti-Beclin-1 antibody (1∶1000, Santa Cruz, CA, USA) and polyclonal rabbit anti-p62/SQSTSM1 antibody (1∶500, Abgent, CA, USA) were used for immunoreactivity detection.

Techniques: Western Blot, Expressing, Quantitation Assay

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: A. EAC cell lines OE19 and OE33 were treated with increasing concentrations of the autophagy inhibitor chloroquine (CQ) for 48hr followed by immunoblotting of the autophagy markers p62 and LC3B. Total protein was visualized as loading control. Representative blot is shown. B. Quantification of triplicate repetitions of experiments shown in A.

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques: Western Blot, Control

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: EAC tissue exhibiting A. low p62 cytoplasmic staining, B. high p62 cytoplasmic staining C. both high p62 dot-like and diffuse cytoplasmic staining, and D. high p62 nuclear positivity. Representative images were taken on a Zeiss Axioskop microscope at 40X objective magnification and corrected for brightness. Insert in A. and C. is for better visualization of the hardly visible dots.

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques: Staining, Microscopy

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: Summary of staining patterns

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques: Staining

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: p62 dot-like, p62 cytoplasmic and p62 nuclear staining patterns correlated to clinicopathological features respectively

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques: Staining

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: p62 sum score and clinicopathological features

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques:

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: A. Dot-like, B. cytoplasmic or C. nuclear staining classified as either low or high. D. p62 sum score. For each curve the p-value is displayed on the bottom right-hand corner.

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques: Staining

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: A. Kaplan-Meier survival curve analysis using classification strategy based on subcellular location and expression of p62. The cohort was subdivided into three groups based on their p62 cytoplasmic (including dot-like) and nuclear staining patterns: low p62 cytoplasmic/low p62 nuclear (LL), either low p62 cytoplasmic/high p62 nuclear or vice versa (mixed), high p62 cytoplasmic/high p62 nuclear (HH). B. Kaplan-Meier survival curve analysis using classification strategy based on LC3B dot-like expression and total p62 expression. The cohort was subdivided into three groups: low LC3B/low p62 (LL), either low LC3B/high p62 or vice versa (mixed), high LC3B/high p62 (HH). For each curve the p-value is displayed on the bottom right-hand corner.

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques: Expressing, Staining

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: p62 cytoplasmic/p62 nuclear classification was shown to be an independent prognostic parameter in a multivariate analysis encompassing pT category, pN category, absence or presence of distant metastasis, lymphatic invasion and perineural invasion as well as grading

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques:

Journal: Oncotarget

Article Title: Prognostic relevance of autophagy markers LC3B and p62 in esophageal adenocarcinomas

doi: 10.18632/oncotarget.9649

Figure Lengend Snippet: p62 cytoplasmic/p62 nuclear classification was shown to be an independent prognostic parameter in a multivariate analysis encompassing UICC and grading

Article Snippet: The anti-p62/SQSTM1 antibody (MBL rabbit polyclonal, #PM045, LabForce, Nunningen, Switzerland) was diluted 1:9000 in tris buffer and incubated at 95°C for 30 min. Visualization was performed using the Bond Polymer Refine Detection kit (Leica Biosystems, product number) per manufacturer's instructions.

Techniques:

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