Journal: Clinical and Experimental Hepatology

Article Title: Comparison of growth features and cancer stem cell prevalence in intrahepatic and extrahepatic cholangiocarcinoma cell lines

doi: 10.5114/ceh.2022.114192

Figure Lengend Snippet: Expression of stem cell surface markers (SCSMs) in cancer stem cell (CSC) in I-CCA (CCLP-1, HuCCT1, SG231), E-CCA (HuH28, KMBC, TFK-1) and nonmalignant H69 cholangiocyte cell lines. Cells were stained with isotype-matched mABs (IgG1) or fluorescence-conjugated mAbs against CD13, CD24, CD44, CD90, CD133, and EpCAM and analyzed by flow cytometry. A ) Cells stained with IgG1 (control/red color) and mAbs against the SCSM (blue color). B ) Prevalence of SCSM positive population in each cell line. Data presented as the mean ±SD from triplicate experiments

Article Snippet: Cells were then transferred into 75 mm polystyrene round-bottom test tubes (BD Falcon, USA) at a concentration of 1 × 10 6 cells/ml, stained with fluorescence-conjugated antibodies (anti-CD13-APC, anti-CD24-APC-eFlour 780, anti-CD44-eFluor 450, anti-CD133-PE, anti-EpCAM-FITC, and 7-AAD) as previously described [ ].

Techniques: Expressing, Staining, Fluorescence, Flow Cytometry, Control

Journal: Cellular immunology

Article Title: Multiple genetic programs contribute to CD4 T cell memory differentiation and longevity by maintaining T cell quiescence

doi: 10.1016/j.cellimm.2020.104210

Figure Lengend Snippet: a) Schematics of the experimental design. 6000 DO11.10 Tg cells, specific to the OVA (323–339) epitope in complex with H-2 d , were adoptively transferred into 8-week old BALB/c recipient mice 24 h before immunization with Vaccinia-OVA virus. Dynamics of gene expression were studied in FACS-sorted CD4 KJ1.26 + CD44 + T cells from lymph nodes and spleens of the recipient mice at day 9, 5 weeks (35 days), 6 months (180 days) and 10.5 months (320 days) post immunization. FACS-sorted CD4 KJ1.26 + CD44 − T cells from age-matched naïve mice served as age control for day 0 and 10.5 months in microarray experiments. For each time point, cell samples were pooled from 5 mice for each cage with 3 cages in total, thus 3 biological replicates were used for statistics in microarray. b) . Selection of gene clusters based on expression dynamics. The microarray data was submitted into Expander software, which grouped all the genes into 16 clusters based on their expression patterns on day 0, day 9, week 5, month 6, and month 10.5 post immunization. The y-axis shown in each cluster is the fold changes of the relative intensities of genes normalized by log2.

Article Snippet: Mouse FITC-CD4 and eFluor 605-CD44 [BD-Pharmingen]; KJ1.26 anti-TCR (Caltag Laboratories), Brilliant Violet 570-CD4 and Alexa Fluor 700-CD44 were from [Biolegend], FITC-Klotho,PE-Cy7-CD99, PE-Cy5-VDR, and PE-Cy7-CNR2 were from [Bioss]; APC-CCR10 and APC-ITGA3 were from [R&D Systems]; Fixable viability dye eFluor 780 was from [eBioscience].

Techniques: Expressing, Microarray, Selection, Software

Journal: Cellular immunology

Article Title: Multiple genetic programs contribute to CD4 T cell memory differentiation and longevity by maintaining T cell quiescence

doi: 10.1016/j.cellimm.2020.104210

Figure Lengend Snippet: a) DR1 mice were injected with H5N1 vaccine and CpG i.p., and 4 months later, draining lymph nodes were extracted and cells incubated for 7 days without any stimulation, or stimulated by either H5N1 HA1 protein or H5N1 vaccine in the media. The cells were then stained with H5N1 ( Left top panels ) or CLIP tetramers ( Left bottom panels ), as well as CD44 and other antibodies for flow cytometry analysis. Dots show the CD44 hi Tetramer + lymphocytes after exclusion of macrophages, B cells and CD8 + T cells. The flow data from 4 repeated experiments is summarized on the right panel , p = 0.88. b) Expression dynamics of several selected membrane associated genes during memory CD4 T cell development ( c ) Expression of different selected membrane/surface markers on CD4 CD44 hi H5N1 tetramer + memory cells in spleen ( orange ), CD4 CD44 hi H5N1 tetramer + activated cells ( red ) and CD4 CD44 lo naïve cells ( blue ). The cells are from the CD4 CD44 hi Tetramer + population (for activated CD4 T cells and CD4 memory T cells) or CD4CD44 lo population (for naïve CD4 T cells) shown in . d) Summary of MFI levels in protein expression among memory CD4 T cells, activated CD4 T cells and naïve T cells from 3 repeated experiments. Data are presented as mean ± standard deviation. Asterisks denote statistical significance: **** P < .0001, *** P < .001, ** P < .01, * P < .05, Student’s two-tailed paired ratio T-test.

Article Snippet: Mouse FITC-CD4 and eFluor 605-CD44 [BD-Pharmingen]; KJ1.26 anti-TCR (Caltag Laboratories), Brilliant Violet 570-CD4 and Alexa Fluor 700-CD44 were from [Biolegend], FITC-Klotho,PE-Cy7-CD99, PE-Cy5-VDR, and PE-Cy7-CNR2 were from [Bioss]; APC-CCR10 and APC-ITGA3 were from [R&D Systems]; Fixable viability dye eFluor 780 was from [eBioscience].

Techniques: Injection, Incubation, Staining, Flow Cytometry, Expressing, Standard Deviation, Two Tailed Test