Journal: Scientific reports

Article Title: miRNA in situ hybridization in circulating tumor cells--MishCTC.

doi: 10.1038/srep09207

Figure Lengend Snippet: Figure 1 | Schematic illustration of the MishCTC method for simultaneous miRNA and CK detection via immunocytochemistry. (A) Recovery of peripheral blood into an EDTA tube; (B) blood transfer into a density-gradient centrifuge tube; (C) centrifugation at 700 3 g for 30 min; (D) recovery of the interphase layer, which contains mononuclear and tumor cells, and immunomagnetic labeling with magnetic microbeads conjugated to an anti-CK antibody; (E) magnetic cell separation using a MiniMACS separator and a pre-filled separation column; (F) elution of retained cells; (G) application of the cells to a polylysine glass slide using CytoSpin centrifugation; and (H) MishCTC detection of miRNA and CK. Mr. Juan M. Agudo helped to prepare this artwork.

Article Snippet: The samples were then incubated for 15 min in a blocking solution (0.1% Tween, 2% sheep serum and 1% BSA in 13 PBS) followed by incubation for 15 min in a solution containing both a FITC-anti-cytokeratin antibody (clone: CK3-6H5; Miltenyi Biotec) and an anti-DIG alkaline phosphatase antibody (Roche Diagnostics, Germany).

Techniques: Immunocytochemistry, Gradient Centrifugation, Centrifugation, Labeling, Magnetic Cell Separation

Journal: Scientific reports

Article Title: miRNA in situ hybridization in circulating tumor cells--MishCTC.

doi: 10.1038/srep09207

Figure Lengend Snippet: Figure 2 | Image galleries obtained with the MishCTC method. (a) CK and miRNA-21 expression in an MDA-MB468 cell that was spiked into a blood sample from a healthy volunteer. Detection of cytokeratin-positive (CK1) cells (green channel), miRNA-21-positive cells (red channel) and nuclei (blue channel). Epithelial cells were identified in a leukocyte population that did not express miRNA-21. (b) CK and miRNA-21 expression in a CTC from a patient with metastatic lung cancer. All the CTCs that were found within this set of patients were both CK- and miRNA-positive (upper panel). CK expression in a circulating epithelial cell from a cancer-free patient undergoing a nephrectomy. CK protein expression (green) was detected by immunofluorescence, but miRNA-21 could not be detected by in situ hybridization (lower panel).

Article Snippet: The samples were then incubated for 15 min in a blocking solution (0.1% Tween, 2% sheep serum and 1% BSA in 13 PBS) followed by incubation for 15 min in a solution containing both a FITC-anti-cytokeratin antibody (clone: CK3-6H5; Miltenyi Biotec) and an anti-DIG alkaline phosphatase antibody (Roche Diagnostics, Germany).

Techniques: Expressing, Immunofluorescence, In Situ Hybridization

Journal: International journal of molecular sciences

Article Title: Resveratrol Inhibition of the WNT/β-Catenin Pathway following Discogenic Low Back Pain.

doi: 10.3390/ijms23084092

Figure Lengend Snippet: Figure 2. Resveratrol administration restored notochordal cells and cartilage markers. Immunohisto- chemical analysis of KRT18: Sham (A), Control (B), CFA (C), CFA + Resveratrol (D); Immunohisto- chemical analysis of KRT19: Sham (E), Control (F), CFA (G), CFA + Resveratrol (H); Western Blot analysis of Aggrecan (I). * p < 0.05 vs. CFA, ### p < 0.001 vs. sham, *** p < 0.001 vs. CFA. Error bar shown as SEM.

Article Snippet: The sections were then incubated overnight with KRT18 (CSB-MA000256, Cusabio Technology, Houston, TX, USA) or KRT19 (CSB-MA000207, Cusabio Technology).

Techniques: Control, Western Blot