vim sc6260  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: A Keratin 7 and E-Cadherin Signature Is Highly Predictive of Tubo-Ovarian High-Grade Serous Carcinoma Prognosis

doi: 10.3390/ijms22105325

Figure Lengend Snippet: HGSC cell lines and tissues display a high variability in epithelial marker expression. ( A ) E-CADH, KRT7, KRT18, KRT19, and VIM protein expression was observed by Western blot in 18 HGSC cell lines. β-actin was used as normalization control. ( B ) KRT7, KRT18, KRT19, E-CADH, and VIM protein expression was observed by immunofluorescence in a TMA comprised of 101 HGSC tissues. Mean fluorescence intensity (MFI) of markers was quantified in the epithelium following digital image analysis. MFI were categorized into 4 groups of expression: 0 (negative), 1 (low), 2 (moderate), and 3 (high), using 25th, 50th and 75th percentiles, respectively, as cut-offs between groups. Spearman Rho correlations between marker expression are indicated in the table. M: Mesenchymal-like phenotype, E: Epithelial-like phenotype. ( C ) Spearman Rho correlations between KRT7, KRT18, KRT19, E-CADH, VIM, and EMT markers were obtained from protein expression measured by the CPTAC (mass spectrometry) on 274 HGSC cases extracted from TCGA (Ovarian Serous Cystadenocarcinoma dataset). Epithelial markers are highlighted in light grey and mesenchymal markers are highlighted in dark grey. Significant correlations are shown in bold ( p < 0.05).

Article Snippet: Antigen retrieval was performed using Cell Conditioning 1 (Tris-EDTA buffer, pH 7.8, for KRT7, KRT18, KRT19, and VIM) or 2 (citrate buffer, pH 6.0, for E-CADH) (Ventana Medical Systems) and slides were then incubated for one hour with the primary mouse monoclonal antibodies against E-CADH (1/100, G10 sc-8426, Santa Cruz Biotechnology, Dallas, TX, USA), KRT7 (1/200, CLONE OV-TL 12/30, Thermo Scientific, Waltham, MA, USA), KRT18 (1/200, sc-6259, Santa Cruz Biotechnology), KRT19 (1/200, MS-198-P, Thermo Scientific) or VIM (1/200, sc6260, Santa Cruz Biotechnology).

Techniques: Marker, Expressing, Western Blot, Control, Immunofluorescence, Fluorescence, Mass Spectrometry

Journal: International Journal of Molecular Sciences

Article Title: A Keratin 7 and E-Cadherin Signature Is Highly Predictive of Tubo-Ovarian High-Grade Serous Carcinoma Prognosis

doi: 10.3390/ijms22105325

Figure Lengend Snippet: KRT7, KRT19, and E-CADH expression associate with prognosis in the HGSC Discovery cohort ( n = 101). ( A – E ) Kaplan–Meier curves of E-CADH ( A ), KRT7 ( B ), KRT18 ( C ), KRT19 ( D ), and VIM ( E ) expression associations with progression-free survival (left) and overall survival (right). MFI measured expression of markers was dichotomized by the median for KRT18 and VIM and by the 75th percentile for E-CADH, KRT7, and KRT19, into groups of low and high expression. p values are indicated, ** = p < 0.01. Number of patients and estimated median survival months (mo) are indicated for each group.

Article Snippet: Antigen retrieval was performed using Cell Conditioning 1 (Tris-EDTA buffer, pH 7.8, for KRT7, KRT18, KRT19, and VIM) or 2 (citrate buffer, pH 6.0, for E-CADH) (Ventana Medical Systems) and slides were then incubated for one hour with the primary mouse monoclonal antibodies against E-CADH (1/100, G10 sc-8426, Santa Cruz Biotechnology, Dallas, TX, USA), KRT7 (1/200, CLONE OV-TL 12/30, Thermo Scientific, Waltham, MA, USA), KRT18 (1/200, sc-6259, Santa Cruz Biotechnology), KRT19 (1/200, MS-198-P, Thermo Scientific) or VIM (1/200, sc6260, Santa Cruz Biotechnology).

Techniques: Expressing