Journal: Cells

Article Title: Neuroprotective and Anti-Inflammatory Effects of Linoleic Acid in Models of Parkinson’s Disease: The Implication of Lipid Droplets and Lipophagy

doi: 10.3390/cells11152297

Figure Lengend Snippet: Effect of LA on lipophagic flux and LD dynamics in the SH-SY5Y PD model. The human cell line SH-SY5Y was exposed to 35 µM 6-OHDA in the presence or not of LA, PF, and CQ. ( A ) Representative maximum intensity projection of confocal images showing the immunocytochemical analysis with the BODIPY 493/503 marker (green), ADRP (red), and p62 (magenta) on SH-SY5Y cells. ( B ) Colocalization analysis of BODIPY 493/503 on ADRP with the Manders overlap coefficient. ( C ) Analysis of BODIPY 492/503 particles per cell. ( D ) Colocalization analysis of BODIPY 493/503 on p62 with the Manders overlap coefficient. ( E ) Analysis of BODIPY 492/503 particles per cell. ( F ) Analysis of the size of BODIPY 492/503 particles per cell. ( G ) Analysis of p62 punctae per cell. ( H ) Colocalization analysis of p62 on BODIPY 493/503 with the Manders overlap coefficient. Graphs show the mean ± SD of 6 independent experiments per group. Data were analyzed by one-way analysis of variance (ANOVA), followed by a multiple comparison test with Bonferroni in some cases and one-way analysis of variance (ANOVA) followed by the Games–Howell correction in other cases. *** p < 0.001; * p < 0.05 vs. control or between two specific groups when indicated). ( I ) Determination of the BODIPY 493/503 signal by flow cytometry. The mean BODIPY 493/503 signal normalized by the control for the different treatment groups is shown. The results correspond to 6 independent experiments per group. Data were analyzed using the Kruskal–Wallis test followed by Dunn’s multiple comparisons test. * p < 0.05 vs. control or between two specific groups when indicated.

Article Snippet: The following antibodies were used: α-cleaved caspase 3 (1:500, R&D Bio-Tech., Abingdon, UK, AF835), α-COX2 (1:200, Santa Cruz Biotechnology, Heidelberg, Germany, sc-376861), α-SQSTM1 (p62, 1:300, Cusabio, Houston, TX, USA, CSB-PA615696LA01HU), and/or α-ADRP (1:200, Santa Cruz, sc-377429).

Techniques: Marker, Comparison, Control, Flow Cytometry

Journal: Cells

Article Title: Neuroprotective and Anti-Inflammatory Effects of Linoleic Acid in Models of Parkinson’s Disease: The Implication of Lipid Droplets and Lipophagy

doi: 10.3390/cells11152297

Figure Lengend Snippet: Study of the LA effect on lipophagic flux and LDs dynamics on SH-SY5Y PD model. Representative maximum intensity projection of confocal images with the BODIPY 493/503 marker (green), ADRP (red), and p62 (magenta) from the SH-SY5Y cell line treated or not with LA, 6-OHDA, PF ( A ), and/or CQ ( B ) are shown.

Article Snippet: The following antibodies were used: α-cleaved caspase 3 (1:500, R&D Bio-Tech., Abingdon, UK, AF835), α-COX2 (1:200, Santa Cruz Biotechnology, Heidelberg, Germany, sc-376861), α-SQSTM1 (p62, 1:300, Cusabio, Houston, TX, USA, CSB-PA615696LA01HU), and/or α-ADRP (1:200, Santa Cruz, sc-377429).

Techniques: Marker

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: