Journal: Scientific Reports

Article Title: Fast and efficient microfluidic cell filter for isolation of circulating tumor cells from unprocessed whole blood of colorectal cancer patients

doi: 10.1038/s41598-019-44401-1

Figure Lengend Snippet: Microscopy images showing cells retained at a central region of the CROSS chip ( A–E ) and identified by CellSearch ( F ). Isolated cells trapped between pillars of the CROSS chip were stained with the following antibodies: anti-pan Cytokeratin-FITC, anti-CD45-Cy5 and anti-Vimentin-eFluor 570, and the nuclear marker DAPI ( A – E ). In the case of CellSearch, staining was done with anti-Cytokeratins 8, 9, 18-PE, anti-CD45-APC and DAPI ( F ). Overlay of the fluorescence microscopy images is shown in color ( A – F ). Different cell populations with distinct expression profiles can be observed: Epithelial CTCs ( A,B ), EMT/MET CTCs ( C ), WBCs ( D ), and CTC clusters ( E ).

Article Snippet: Isolated cells from patient samples were permeabilized with 0.25% Triton X-100 solution (Sigma Aldrich, USA) and fluorescently labeled inside the microfluidic device with anti-pan CK-FITC (clone C-11, recognizes human cytokeratins 4,5,6,8,10,13, and 18, Sigma; 1:100 in PBS-2% BSA), anti-Vimentin eFluor 570 (eBioscience, 1:50 in PBS-2%BSA) and anti-CD45-Cy5 (Abcam; 1:25 in PBS-2%BSA) antibodies for 1 h. DAPI (Sigma Aldrich, 1:1000 in PBS-2%BSA) was used as a nuclear marker.

Techniques: Microscopy, Isolation, Staining, Marker, Fluorescence, Expressing

Journal: International Journal of Cancer. Journal International du Cancer

Article Title: SPARC-induced increase in glioma matrix and decrease in vascularity are associated with reduced VEGF expression and secretion

doi: 10.1002/ijc.23450

Figure Lengend Snippet: Matrix assessment in control- (−SPARC) and SPARC- (+SPARC) expressing tumors. ( a ) The level of SPARC in the control and SPARC-expressing cells was assessed by Western blot analysis prior to brain implantation. P-parental clonal U87MG-derived cell line, VC1 and VC2-vector control cell lines 1 and 2, S1 and S2-SPARC-transfected cell lines 1 and 2. +C-SPARC protein positive control. L: lysate, M: medium. (∧) indicates that the signal is on the same gel but moved closer. Actin detection on the same lysate blot was used as a loading control. ( b ) Representative images of tumor sections either immunohistochemically stained for SPARC (×40) or procollagen I (Pro-CN I; ×20), or enzyme-treated or untreated sections stained with Schiff's reagent (PAS ± diastase; ×40) or stained with picrosirius red and imaged under polarized light (×20).

Article Snippet: Immunohistochemistry was performed using the Biocare Medical Nemesis 7200 (Concord, CA) stainer and reagents for anti-SPARC antibody (1:13,333 dilution in 0.25% BSA in PBS), and anti-procollagen I antibody (1:250 dilution), anti-factor VIII (1:1,200), anti-VEGF antibody (#SC-7269, 1:250) or anti-VEGFR2 antibody (1:3,000) were all diluted in DaVinci antibody diluent.

Techniques: Expressing, Western Blot, Derivative Assay, Plasmid Preparation, Transfection, Positive Control, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: No apparent p53 activation in CRISPR‐engineered gene‐edited rabbits

doi: 10.1111/jcmm.16960

Figure Lengend Snippet: Summary of the CRISPR induced a p53 expression changes in rabbits

Article Snippet: After blocking with 10% goat serum in PBS for 1 h at room temperature and incubated with an antibody against p53 (diluted in PBS with 1% BSA) at 4°C overnight (60283–2, Proteintech) was used.

Techniques: CRISPR, Expressing, Mutagenesis

Journal: Journal of Cellular and Molecular Medicine

Article Title: No apparent p53 activation in CRISPR‐engineered gene‐edited rabbits

doi: 10.1111/jcmm.16960

Figure Lengend Snippet: Expression of p53 not changes in rabbits generated by Cas9 or BE‐edited. When CRISPR/Cas9 make a single DSB in chromosome, inaccurately repair via nonhomologous end joining (NHEJ), this is the normal genome‐editing outcome, which not induces the expression of p53 changes in rabbits. On the contrary, the BEs‐mediated gene‐editing is not generated the DSB, it only provides precise editing at a single base‐pair conversion not causes the p53 changes

Article Snippet: After blocking with 10% goat serum in PBS for 1 h at room temperature and incubated with an antibody against p53 (diluted in PBS with 1% BSA) at 4°C overnight (60283–2, Proteintech) was used.

Techniques: Expressing, Generated, CRISPR