Journal: eLife

Article Title: A concerted mechanism involving ACAT and SREBPs by which oxysterols deplete accessible cholesterol to restrict microbial infection

doi: 10.7554/eLife.83534

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Article Snippet: Antibody , anti-ACAT1 (Rabbit Polyclonal) , Novus , CatNB400-141; RRID: AB_10001588 , WB (1:1,000).

Techniques: Transfection, Construct, Flow Cytometry, Recombinant, Sequencing, Bicinchoninic Acid Protein Assay, Microscopy, Protease Inhibitor, Modification, Saline, Methylation, Software, CRISPR, Imaging

Journal: Scientific reports

Article Title: Combined microfluidic enrichment and staining workflow for single-cell analysis of circulating tumor cells in metastatic prostate cancer patients.

doi: 10.1038/s41598-024-68336-4

Figure Lengend Snippet: Figure 1. In-cassette staining method for CTC detection. (A) Schematic overview of the workflow. Abbreviations: mononuclear cells (MNC), copy-number alterations (CNA) (B) Recovery of 100 PC3 cells spiked into 10 mL healthy donor blood. The samples were processed with DPBS or DPBS + 1% BSA + 1 mM EDTA buffer on the Parsortix. CKPos, DAPIPos and CD45Neg cells were manually counted as CTCs, and the recovery was calculated from the number of spiked cells vs. number of detected cells. Data is presented as mean ± sd. The comparison between the two groups was done using a Mann–Whitney U test (p = 0.019) (C) Test of two different priming and separation buffers: DPBS or DPBS + 1% BSA + 1 mM EDTA. The harvested cells were plated on microscopy slides and the total number of DAPIPos cells were automatically counted. The harvested cell fraction was calculated from the total number of cells (harvested fraction + cells left in the cassette). Data is presented as mean ± sd. Statistical analysis was performed using Mann–Whitney U test (p = 0.017) (D) Spike in of 100 prostate cancer cells (PC3, LnCAP, DU145, C4-2) as described in B (E) Recovery of 10 PC3 cells spiked into 10 mL healthy donor blood (F) Stability of 100 PC3 cells spiked into 10 mL of healthy blood (CellRescue). The blood was processed after ~ 1 h, 24 h (p = 0.062) or 72 h (p = 0.024) and stained as described in B. The comparison between the three groups was performed using a Mann–Whitney U test (G) Recovery of 100 PC3 cells spiked into a K2EDTA blood tube and processed after 1 h. Data is presented as mean ± sd. (H) Staining of four prostate cancer cell lines (PC3, LNCaP, DU145 and C4-2) with DAPI and anti-CK. The cells were images by imaging flow cytometry (ImageStream).

Article Snippet: The captured cells in the cassette were incubated with permeabilization buffer (20 min) containing separation buffer + 0.15% saponin (Sigma-Aldrich, 47036) followed by immunostaining (90 min) with antibodies against cytokeratin (FITC-conjugated, Macs Miltenyi biotech, 130-118-964, clone CK36H5, 1:100), CD45 (Alexa Fluor 647-conjugated, eBioscience, 51-0459-42, clone HI30, 1:100) and DAPI (Fischer Scientific, D1304, 1:10.000) diluted in separation buffer containing 0.05% saponin.

Techniques: Staining, Comparison, MANN-WHITNEY, Microscopy, Imaging, Flow Cytometry

Journal: iScience

Article Title: Characterizing replisome disassembly in human cells

doi: 10.1016/j.isci.2024.110260

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Article Snippet: Mouse monoclonal anti-MCM7 (clone 141.2) , Santa Cruz , Cat#Sc-9966; RRID: AB_627235.

Techniques: Recombinant, Transfection, Imaging, Isolation, Immunoprecipitation, Sequencing, shRNA, Plasmid Preparation, CRISPR, Marker, Software, Cell Analysis