plectin antibody (plec)  (Thermo Fisher)

Journal: Cancers

Article Title: Plakin Expression in Serous Epithelial Ovarian Cancer Has the Potential to Impede Metastatic Spread and Epithelial-Mesenchymal Transition: A Comparative Expression Analysis of Immunohistochemical and In Silico Datasets.

doi: 10.3390/cancers16234087

Figure Lengend Snippet: Figure 2. Representative images of plakin expression in benign and cancerous solid ovarian tumours evaluated by immunohistochemistry. Representative images of protein expression using immuno- histochemistry in benign ovarian tissue, borderline, Type I and Type II tumours. (a) PPL, (b) PLEC, (c) EVPL immunostaining, and (d) IHC negative control image using secondary-only anti-mouse HRP-labelled DAB staining. Images were taken using a Leica DLMB microscope with attached Leica DFC450C camera and LAS software 4.8.0, ×40 magnification (D ×20 magnification) or from Aperio Imagescope software 12.3.

Article Snippet: The staining process included antigen retrieval at pH 8.5, blocking of endogenous peroxidase activity, and incubating the tissue in primary antibody for 60 min. Each primary mouse anti-human antibody against PPL (G-1:sc-365530), EVPL (F-4:sc-137033), and PLEC (10F6:sc-33649) was used at a 1:100 dilution, was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Immunohistochemistry, Immunostaining, Negative Control, Staining, Microscopy, Software

Journal: Cancers

Article Title: Plakin Expression in Serous Epithelial Ovarian Cancer Has the Potential to Impede Metastatic Spread and Epithelial-Mesenchymal Transition: A Comparative Expression Analysis of Immunohistochemical and In Silico Datasets.

doi: 10.3390/cancers16234087

Figure Lengend Snippet: Figure 3. Quantitative analysis of plakins staining in benign ovarian tissues and serous ovarian tumours. Quantification of plakin protein expression staining using DAB immunohistochemistry. Positivity output from Aperio was normalized to background (non-epithelial cell) staining. Quantifi- cation of IHC staining: (a) PPL, (b) PLEC and (c) EVPL for each of cases sorted by WHO tumour Type, cases sorted by FIGO surgical stage, (blue data points represent WHO Type I cases) and cases sorted by Silverberg–Shimizu tumour grade. For PPL expression at a), statistical significance measured by one-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.005. For PLEC at (b), statistical significance measured by one-way ANOVA, * p < 0.05. For EVPL at (c), statistical significance was measured by one-way ANOVA; no significance was determined between the groups.

Article Snippet: The staining process included antigen retrieval at pH 8.5, blocking of endogenous peroxidase activity, and incubating the tissue in primary antibody for 60 min. Each primary mouse anti-human antibody against PPL (G-1:sc-365530), EVPL (F-4:sc-137033), and PLEC (10F6:sc-33649) was used at a 1:100 dilution, was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Staining, Expressing, Immunohistochemistry

Journal: Cancers

Article Title: Plakin Expression in Serous Epithelial Ovarian Cancer Has the Potential to Impede Metastatic Spread and Epithelial-Mesenchymal Transition: A Comparative Expression Analysis of Immunohistochemical and In Silico Datasets.

doi: 10.3390/cancers16234087

Figure Lengend Snippet: Figure 7. Correlation of PLEC and PPL with EMT markers in WHO Type II primary tumour samples. Pearson correlation analysis of PLEC and PPL staining with EMT marker expression in Type II tumours from IHC staining. Top row: (a) PLEC vs. VIM, (b) PLEC vs. ECAD * p < 0.05, (c) PLEC vs. NCAD, (d) high PLEC (cases above mean PLEC expression) vs. NCAD * p < 0.05. Bottom row: (e) PPL vs. VIM, (f) PPL vs. ECAD, (g) PPL vs. NCAD, (h) high PPL (cases above mean PPL expression) vs. NCAD * p < 0.05.

Article Snippet: The staining process included antigen retrieval at pH 8.5, blocking of endogenous peroxidase activity, and incubating the tissue in primary antibody for 60 min. Each primary mouse anti-human antibody against PPL (G-1:sc-365530), EVPL (F-4:sc-137033), and PLEC (10F6:sc-33649) was used at a 1:100 dilution, was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Staining, Marker, Expressing, Immunohistochemistry

Journal: Cancers

Article Title: Plakin Expression in Serous Epithelial Ovarian Cancer Has the Potential to Impede Metastatic Spread and Epithelial-Mesenchymal Transition: A Comparative Expression Analysis of Immunohistochemical and In Silico Datasets.

doi: 10.3390/cancers16234087

Figure Lengend Snippet: Figure 17. Kaplan–Meier analysis of overall and progression-free survival of patients in relation to mRNA expression of plakins. Cases analysed using the Kaplan–Meier Plotter website, dataset titled Ovarian Serous Cystadenocarcinoma (TCGA, Nature 2011) [49], with these conditions: WHO Type II serous ovarian cancer with p53 mutations, all treatment Types, and auto-threshold selection of high versus low expression. (a) PPL mRNA expression in relation to overall survival, (b) PPL mRNA expression in relation to progression-free survival, (c) PLEC mRNA expression in relation to overall survival, (d) PLEC mRNA expression in relation to progression-free survival, (e) EVPL mRNA expression in relation to overall survival, (f) EVPL mRNA expression in relation to progression- free survival. Sample numbers: n = 418 for OS, n = 398 for PFS, samples without relevant data were omitted.

Article Snippet: The staining process included antigen retrieval at pH 8.5, blocking of endogenous peroxidase activity, and incubating the tissue in primary antibody for 60 min. Each primary mouse anti-human antibody against PPL (G-1:sc-365530), EVPL (F-4:sc-137033), and PLEC (10F6:sc-33649) was used at a 1:100 dilution, was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Selection

Journal: Cancers

Article Title: Plakin Expression in Serous Epithelial Ovarian Cancer Has the Potential to Impede Metastatic Spread and Epithelial-Mesenchymal Transition: A Comparative Expression Analysis of Immunohistochemical and In Silico Datasets.

doi: 10.3390/cancers16234087

Figure Lengend Snippet: Figure 19. Overall survival in relation to high and low expression of PPL (a), PLEC (b), and EVPL (c) in RWH cases (WHO Type II only). Data analysis and survival graphs produced with GraphPad Prism 10.2.0 software of the protein expression and survival data of the WHO Type II cases presented in Figures 1–7. Cases separated by level of plakin protein expression, high—above mean identified in Figure 2, low—below mean in Figure 3. No significant difference was found using Mantel–Cox (log-rank) test.

Article Snippet: The staining process included antigen retrieval at pH 8.5, blocking of endogenous peroxidase activity, and incubating the tissue in primary antibody for 60 min. Each primary mouse anti-human antibody against PPL (G-1:sc-365530), EVPL (F-4:sc-137033), and PLEC (10F6:sc-33649) was used at a 1:100 dilution, was obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Produced, Software

Journal: Scientific Reports

Article Title: Post-translational modifications to hemidesmosomes in human airway epithelial cells following diacetyl exposure

doi: 10.1038/s41598-022-14019-x

Figure Lengend Snippet: Changes in plectin abundance after DA exposure. ( A ) Representative western blot of plectin (PLEC) from epithelial cell culture homogenates at Days 1, 2 and 4 after DA exposure (5.7 and 8.6 mM). ( B ) Semi-quantification of PLEC abundance normalized to lane total protein that decreased significantly at Day 4 in cultures exposed to 8.6 mM DA compared to controls (ANOVA, *p < 0.05; n = 4/group).

Article Snippet: Ten micrograms (μg) total protein were resolved in stain-free, pre-cast 4–15% gradient Tris–Glycine gel (Bio-Rad, Hercules, CA), and stained with primary antibodies of keratin 5 (KRT5; 1:2000, Biolegend, San Diego, CA), integrin beta 4 (ITGβ4, CD104; 1:1000, Invitrogen, Waltham, MA), and plectin (PLEC; 1:1000, ThermoFisher, Waltham, MA).

Techniques: Western Blot, Cell Culture