Journal: Disease Markers

Article Title: EV-Associated MMP9 in High-Grade Serous Ovarian Cancer Is Preferentially Localized to Annexin V-Binding EVs

doi: 10.1155/2017/9653194

Figure Lengend Snippet: EV isolation from ascites. EVs were isolated from ascites with the ligands AV (lane 1), CTB (lane 2), and STB (lane 3) and analyzed by western blotting. As a negative control, no ligand was added (lane 4). 250 μ l of pooled ascites from 6 (a) or 16 (b) ovarian cancer patients were used as starting material. The tetraspanin proteins CD9 and CD63, the transferrin receptor (CD71), and the cytosolic protein Alix were examined, and β -actin was used as a loading control. The normalized band intensities of the analyzed proteins are given below each band. The molecular weight markers are indicated on the left side of the images in kDa.

Article Snippet: The primary antibodies were mouse antibodies against the human proteins β -actin (C4), β -catenin (E-5), CD71 (H68.4), CD9 (C-4), Alix (1A12), CD59 (H-7), EpCAM (0.N.276) (Santa Cruz Biotechnology Inc., USA), CD63 (H5C6) (BD Biosciences, USA), and FN1-EDA (IST-9) (abcam PLC, UK).

Techniques: Isolation, Western Blot, Negative Control, Control, Molecular Weight

Journal: Disease Markers

Article Title: EV-Associated MMP9 in High-Grade Serous Ovarian Cancer Is Preferentially Localized to Annexin V-Binding EVs

doi: 10.1155/2017/9653194

Figure Lengend Snippet: Cancer proteins in EV isolations. (a) EVs isolated from 250 μ l of pooled ascites from 6 ovarian cancer patients (lanes 1–4) or from 5 patients with cirrhosis (lanes 5–8) were analyzed by western blot. As before the ligands AV, CTB, and STB were used for isolation, no ligand was added as a negative control. The protein cellular fibronectin (FN1-EDA), CD59, β -catenin, and epithelial cell adhesion molecule (EpCAM) were examined, and β -actin was used as loading control. The normalized band intensities of the analyzed proteins are given below each band. (b) EVs isolated from 500 μ l of the same samples as described for (a) were analyzed by zymography. The left part of the image (lanes 1–5) shows the ovarian cancer samples and the right part the cirrhosis samples (lanes 6–10). In lanes 1 and 6, 1 μ l of the pooled ascites samples were analyzed. The molecular weight markers are indicated on the left side of the images in kDa.

Article Snippet: The primary antibodies were mouse antibodies against the human proteins β -actin (C4), β -catenin (E-5), CD71 (H68.4), CD9 (C-4), Alix (1A12), CD59 (H-7), EpCAM (0.N.276) (Santa Cruz Biotechnology Inc., USA), CD63 (H5C6) (BD Biosciences, USA), and FN1-EDA (IST-9) (abcam PLC, UK).

Techniques: Isolation, Western Blot, Negative Control, Control, Zymography, Molecular Weight

Journal: Frontiers in Oncology

Article Title: Establishment, characterization, and drug screening of low-passage patient individual non-small cell lung cancer in vitro models including the rare pleomorphic subentity

doi: 10.3389/fonc.2023.1089681

Figure Lengend Snippet: Antibodies used for flow cytometry.

Article Snippet: LYPD3 , APC , Sino Biological Europe GmbH, Düsseldorf, Germany , 11836-H08H.

Techniques: Cytometry

Journal: Frontiers in Oncology

Article Title: Establishment, characterization, and drug screening of low-passage patient individual non-small cell lung cancer in vitro models including the rare pleomorphic subentity

doi: 10.3389/fonc.2023.1089681

Figure Lengend Snippet: Flow cytometry. Histogram overlays of unstained controls (dotted lines for all three cell lines) and measurements for HROLu55 (green), HROLu22 (blue), and HROBML01 (red) for the epitopes CD326, PD-L1, EGFR, CD26, LYPD3, DSG3, CCD59, CD27, and CD90 are shown.

Article Snippet: LYPD3 , APC , Sino Biological Europe GmbH, Düsseldorf, Germany , 11836-H08H.

Techniques: Flow Cytometry

Journal: Oncology Letters

Article Title: Overexpression of close homolog of L1 enhances the chemosensitivity of lung cancer cells via inhibition of the Akt pathway

doi: 10.3892/ol.2020.11972

Figure Lengend Snippet: CHL1 is downregulated in DDP and PTX-resistant A549 cells. (A) Cell survival of A549 and A549-resistant cells (A549/DDP and A549/PTX) treated with increasing concentrations of DDP and PTX, as assessed by MTT assay. (B) The IC 50 values of DDP in A549/DDP and A549 cells, and the IC 50 values of PTX in A549/PTX and A549 cells. *P<0.05 vs. A549 cells. (C) Western blotting demonstrated the expression of drug resistance-related proteins MDR1, MRP and LRP in A549 cells and A549-resistant cells (A549/DDP and A549/PTX). *P<0.05 vs. A549 cells. The protein and mRNA expression levels of CHL1 in A549 cells and A549-resistant cells (A549/DDP and A549/PTX) were analysed by (D) western blotting and (E) reverse transcription-quantitative PCR, respectively. *P<0.05 vs. A549 cells. (F) The mRNA expression of CHL1 in H460 and H460/DDP cells in the GSE21656 dataset. *P<0.05 vs. H460 cells. CHL1, close homolog of L1; DDP, cisplatin; PTX, paclitaxel; MDR1, multi-drug resistance gene 1; MRP, multidrug resistance-associated protein; LRP, low-density lipoprotein receptor-related protein; IC50, half maximal inhibitory concentration.

Article Snippet: Next, the proteins were transferred onto a polyvinylidene membrane (Thermo Fisher Scientific, Inc.), blocked with 5% BSA (Thermo Fisher Scientific, Inc.) for 2 h at 4°C, and incubated overnight at 4°C with primary antibodies against CHL1 (1:500; cat. no. 25250-1-AP; ProteinTech, Inc.), multi-drug resistance gene 1 (MDR1; 1:500; cat. no. 22336-1-AP; ProteinTech, Inc.), multidrug resistance-associated protein (MRP; 1:500; cat. no. 67228-1-Ig; ProteinTech, Inc.), low-density lipoprotein receptor-related protein (LRP; 1:500; cat. no. 22336-1-AP; ProteinTech, Inc.), phosphorylated (p)-Akt (1:1,000; cat. no. ab38449; Abcam,) and Akt (1:2,000; cat. no. ab227385; Abcam).

Techniques: MTT Assay, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Concentration Assay