Journal: Nature Communications

Article Title: KK-LC-1 as a therapeutic target to eliminate ALDH + stem cells in triple negative breast cancer

doi: 10.1038/s41467-023-38097-1

Figure Lengend Snippet: a Volcano plot showing up-regulated genes in docetaxel-resistant MDA-MB-231 cells (DTX_R) compared to their parental cells (Parental) (Log2 Fold change > 6, adj P value < 0.05, P values were determined using moderated t -statistic, adj P -value was calculated using Benjamini and Hochberg’s method and tails were two-sided). b Up-regulated genes in docetaxel-resistant MDA-MB-231 cells were intersected with the up-regulated genes in TNBC , . Expression levels of two genes were significantly higher in docetaxel-resistant MDA-MB-231 cells. c Heatmap showing expression of the two highly expressed genes. d Schematic showing the identification and isolation of primary ALDH + and ALDH - cells. e Among the two highly expressed genes, KK-LC-1 had the highest expression in ALDH + cells compared to ALDH − cells and was selected as a potential candidate that may regulate ALDH + cells in TNBC; data are presented as mean ± SD from three biologically independent experiments, statistical significance was determined using Student’s t -test and was two-sided. f, g : KK-LC-1 knockout significantly reduced the proportion of ALDH + cells and ALDH + CD44 high CD24 low cells in MDA-MB-231 cells; data are presented as mean ± SD from three biologically independent experiments and statistical significance was determined using Student’s t -test and was two-sided. h The proportion of ALDH + cells was significantly lower in ALDH + sh- KK-LC-1 #1 cells compared to ALDH + NC cells after 2 weeks in culture; data are presented as mean ± SD from three biologically independent experiments, statistical significance was determined using Student’s t -test and was two-sided. i A significantly higher proportion of ALDH + cells was derived from CD44 high CD24 low KK-LC-1 +/+ MDA-MB-231 compared to CD44 high CD24 low KK-LC-1 −/− MDA-MB-231 cells after 2 weeks in culture; data are presented as mean ± SD from three biologically independent experiments, statistical significance was determined using Student’s t -test and was two-sided. Source data are provided as a Source data file.

Article Snippet: MDA-MB-231 cells and MDA-MB-468 cells were resuspended in PBS and stained with phycoerythrin (PE)-conjugated anti-CD24 at a dilution of 1:100 (Cat: 130-112-656, Miltenyi Biotec) and allophycocyanin (APC)-conjugated anti-CD44 at a dilution of 1:500 (Cat: 12211-MM02-A, SinoBiological) for 30 min at 4 °C.

Techniques: Expressing, Isolation, Knock-Out, Derivative Assay

Journal: Genes & Diseases

Article Title: ROS and Lipid Droplet accumulation induced by high glucose exposure in healthy colon and Colorectal Cancer Stem Cells

doi: 10.1016/j.gendis.2019.09.010

Figure Lengend Snippet: 72 hrs in HG cell culture medium is sufficient to increase CSC markers in CR-CSCs. a) Primary CR-CSC spheroids (#1, #4, #8 and #9) were cultured in ultra-low adhesion flasks and morphology was assessed by phase-contrast microscopy (scale bar: 1000 μm). b) Fold variation of protein expression in CR-CSCs treated with HG versus NG. Data are representative of 3 independent experiments performed with cells derived from 3 different CR-CSC lines. c) Representative Western blot of PI3K, phosphorylated (p-AKT) and total AKT in CR-CSC treated with HG or NG (#9 CR-CSC line is shown). β-Actin has been used as loading control. The uncropped blots are reported in Supplementary Fig. 1 . d) Fold variation of positivity for CD133 or CD44v6 in CR-CSCs treated as in ( b and c ), measured by FACS.

Article Snippet: Cells were dissociated, harvested, washed twice in Phosphate Buffer Solution (PBS) 1X (Thermo Fisher Scientific; #10010023) and stained with conjugated antibodies CD44v6-APC (2F10, mouse IgG1) (R&D systems; #FAB3660A), CD133/2-APC (293C3-APC) (Miltenyi Biotec; #130-090-854), or corresponding isotype-matched control (IMC) CD3e-APC (UCHT1, mouse IgG1) (R&D systems; #FAB100A).

Techniques: Cell Culture, Microscopy, Expressing, Derivative Assay, Western Blot, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Implantation with SHED sheet induced with homogenate protein of spinal cord promotes functional recovery from spinal cord injury in rats

doi: 10.3389/fbioe.2023.1119639

Figure Lengend Snippet: The construction and characteristics of hp-SHED sheet, which possesses a geometric structure similar to that of spinal cord tissue. (A) The surface markers of SHED, including CD44, CD73, CD90, CD105 and HLA were analyzed by flow cytometry. (B) Map of the morphology of the SHED. (C) SHED formed single CFU clusters in culture, the quantitative result was indicated in right panel. Data shown as mean ± SD. (D) Osteogenic differentiation of SHED. Alizarin red staining was performed at day 21 after osteogenic induction. (E) Adipogenic differentiation of SHED. Oil Red O staining was performed at day 28 after adipogenic induction. (F) The proliferation curve of SHED with different concentrations of ascorbic acid (AA) was analyzed by CCK8 assay. (mean ± SD, *** p < 0.001). (G) Construction of hp-SHED sheet. (H) Representative images of hp-SHED sheet sections. (I) Collagens in hp-SHED sheet were characterized by CLSM.

Article Snippet: Under light-proof conditions, 1 μl of antibodies were added to each EP tube, Anti-Human CD73 PE, Anti-Human CD105 (Endoglin) PE, Anti-Human/Mouse CD44 APC, Anti-Human CD90 (Thy-1) PE, Anti-Human HLA-DR FITC (BioGems, United States), control group plus PBS.

Techniques: Flow Cytometry, Staining, CCK-8 Assay

Journal: Cancer research

Article Title: Microenvironment-driven dynamic chromatin changes in glioblastoma recapitulate early neural development at single-cell resolution

doi: 10.1158/0008-5472.CAN-22-2872

Figure Lengend Snippet: RESOURCES TABLE

Article Snippet: For inducible DLX5 expression, the same protocol was followed in GLICO. . Flow Cytometry analysis GSCs were stained with CD44 Antibody coupled to APC (#130–113-338, Miltenyi Biotec) followed manufacturer’s Cell surface flow cytometry staining protocol.

Techniques: Recombinant, Gene Expression, Software