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: