Journal: New biotechnology

Article Title: Clinically compatible advances in blood-derived endothelial progenitor cell isolation and reprogramming for translational applications.

doi: 10.1016/j.nbt.2021.02.001

Figure Lengend Snippet: Fig. 1. EPC morphology and gene expression. (A) Appearance of a freshly isolated EPC colony at passage 0 on day 9 of culture. (B) Appearance of EPCs in culture at passage 2, (C–F) EPCs express the endothelial cell specific markers CD31 (C, red, DAPI blue) and CD144 (D, green, DAPI blue) and pluripotency associated markers NANOG (E, red) and REX-1 (F, red, DAPI blue). Images A-B at 50X magnification, images C, D, E, F at 100X magnification.

Article Snippet: Primary EPCs were stained using specific antibodies against CD31 (Cat. no. BBA7, R&D Systems, Abingdon, UK), CD144 (Cat. no. MAB9381, R&D Systems) and REX1 (Cat. no. AF3598, R&D Systems), or by using ULEX (Cat. no. RL-1062-2, Vector Laboratories) or DAPI (Cat. no. H-1200-10, Vector Laboratories) at the dilution recommended by the manufacturer.

Techniques: Gene Expression, Isolation

Journal: Infection and Immunity

Article Title: Requirement for Intercellular Adhesion Molecule 1 and Caveolae in Invasion of Human Oral Epithelial Cells by Porphyromonas gingivalis

doi: 10.1128/iai.73.10.6290-6298.2005

Figure Lengend Snippet: FIG. 1. Inhibition of P. gingivalis invasion into KB cells by anti-ICAM-1 antibody. The cells were preincubated with or without either goat polyclonal anti-ICAM-1 antibody, goat polyclonal anti-E-cadherin antibody, or goat IgG at 37°C for 30 min as indicated, and then P. gingivalis organisms were incubated with the cells at 37°C for 90 min. Invasion of the cells by P. gingivalis at a multiplicity of infection of 100 was determined by an invasion assay. The assays were carried out in triplicate as described in Materials and Methods. The CFU of P. gingivalis invasion without antibodies was set at 100%. **, significantly different (P 0.01) from the mean value for preincubation without antibody. Error bars indicate standard errors of the means.

Article Snippet: Recombinant human ICAM-1, mouse monoclonal antibody specific for ICAM-1, and goat polyclonal antibody specific for E-cadherin were purchased from R&D Systems Inc. (Minneapolis, Minn.).

Techniques: Inhibition, Incubation, Infection, Invasion Assay

Journal: The Journal of Biological Chemistry

Article Title: N-acetyltransferase 10 promotes glioblastoma malignancy via mRNA stabilization of jumonji and AT-rich interaction domain containing 2

doi: 10.1016/j.jbc.2025.108544

Figure Lengend Snippet: Suppression of GBM stemness by knockout of NAT10 . A , the proliferation of naive and NAT10 knockout (KO) U251 and A172 cells. Values show the mean with S.D. (n = 12). The cell viability of seeding day (day 0) was set at 1.0. ∗∗; p < 0.01 significant difference from naive group at corresponding time points. ( F 9,110 = 69.073 p < 0.001 for U251 cells, F 9,110 = 2331.346 p < 0.001 for A172 cells; ANOVA with the Tukey-Kramer post hoc test). Each upper panel shows the NAT10 protein levels in naive and NAT10 KO U251 and A172 cells. B , the spheroid formation ability of naive and NAT10 KO U251 and A172 cells. Each left panel shows a representative photograph of Hoechst33342-stained spheroids-formed by naive or NAT10 KO U251 or A172 cells. Each right panel shows the number of spheroids and the distribution of their diameters. Values show the mean with S.D. (n = 5 for U251 cells, n = 4 for A172 cells). ∗∗; p < 0.01, significant difference between the two groups ( t = 13.355 for U251 cells, t = 2.663 for A172 cells, Welch’s t test). C , the invasion ability of naive and NAT10 KO U251 and A172 cells. Microphotographs show invasion of cells into 3D collagen gel. D , the sensitivity of naive and NAT10 KO U251 and A172 cells to temozolomide. Naive and NAT10 KO cells were treated with indicated concentration of temozolomide for 96 h. Values show the mean with S.D. (n = 8 for U251 cells, n = 6 for A172 cells). Cell viability of vehicle-treated groups was set at 1.0. ∗∗; p < 0.01, ∗; p < 0.05; significant difference between the two groups at corresponding concentration points. ( F 7 ,56 = 55.596 p < 0.001 for U251 cells, F 7,40 = 7.381 p < 0.001 for A172 cells; ANOVA with the Tukey-Kramer post hoc test). E , protein expression levels in SOX2, OCT4, KLF4 in naive or NAT10 KO U251 cells. Protein levels were normalized to those of p84 expression levels. Values show the mean with S.D. (n = 5–6). ∗∗; p < 0.01, ∗; p < 0.05; significant difference between the two groups ( t 10 = 4.134 for SOX2, t 10 = 3.001 for OCT4, t 8 = 4.491 for KLF4, Student’s t test). F , protein expression levels in N-CADHERIN, VIMENTIN in naive and NAT10 KO U251 cells. Protein levels were normalized to those of β-ACTIN expression levels. Values show the mean with S.D. (n = 6). ∗∗; p < 0.01 significant difference between the two groups ( t 10 = 3.979 for N-CADHERIN, t 10 = 5.635 for VIMENTIN, Student’s t test).

Article Snippet: Membranes were reacted with antibodies against NAT10 (13365-1-AP, Proteintech, Wuhan, China, RRID:AB_2148944), p84 (THOC1, 10920-1-AP, Proteintech, RRID:AB_2202239), SOX2 (AF2018, R&D systems, RRID:AB_355110), KLF4 (#4038, Cell Signaling Technology, RRID:AB_2265207), OCT4 (#4038, Cell Signaling Technology), N-CADHERIN (AF6426, R&D systems, RRID:AB_10718850), VIMENTIN (MAB21052, R&D systems, RRID:AB_2832972), SUZ12 (#3737, Cell Signaling Technology, RRID:AB_2196850), JARID2 (#13594, Cell Signaling Technology, RRID:AB_2798269), THRAP3 (TRAP150, sc-133250, Santa Cruz Biotechnology, RRID:AB_2202901), MAD1L1 (MAD1, 18322-1-AP, Proteintech, RRID:AB_2139251) and β-ACTIN conjugated with horseradish peroxidase (sc-47778, Santa Cruz Biotechnology, RRID:AB_2202239).

Techniques: Knock-Out, Staining, Concentration Assay, Expressing

Journal: The Journal of Biological Chemistry

Article Title: N-acetyltransferase 10 promotes glioblastoma malignancy via mRNA stabilization of jumonji and AT-rich interaction domain containing 2

doi: 10.1016/j.jbc.2025.108544

Figure Lengend Snippet: Contribution of JARID2 to the maintenance of stemness properties of GBM cells . A , decrease in the growth ability of U251 and A172 cells by downregulation of JARID2. Each upper panel show JARID2 protein levels in U251 and A172 cells transfected with shRNA against JARID2 or scramble shRNA (shScramble). Each below graph shows the cell viability of seeding day (day 0) was set at 1.0. Values show the mean with S.D. (n = 6). ∗∗; p < 0.01 significant difference from shScramble group at corresponding time points. ( F 9,50 = 1636.125 p < 0.001 for U251 cells, F 9,50 = 140.977 p < 0.001 for A172 cells, ANOVA with the Tukey-Kramer post hoc test). B , the spheroid formation ability of U251 and A172 cells transfected with shRNA against JARID2 or shScramble. Eash left panel shows a representative photograph of Hoechst33342-stained spheroids formed by shScramble- or shJARID2-transfected U251 and A172 cells. Right panel shows the number of spheroids and the distribution of their diameters. Values show the mean with S.D. (n = 5–6 for U251 cells, n = 8 for A172 cells). ∗∗; p < 0.01 significant difference between the two groups ( t = 11.897. for U251 cells, t = 3.412 for A172 cells, Welch’s t test). C , the invasion ability of U251 and A172 cells transfected with shRNA against JARID2 or shScramble. Microphotographs show invasion of cells into 3D collagen gel. D , the protein expression levels of SOX2, OCT4, and KLF4 in U251 cells transfected with shRNA against JARID2 or scramble shRNA (shScramble). Values of protein levels were normalized to p84 protein levels. Values show the mean with S.D. (n = 5). ∗∗; p < 0.01 significant difference between the two groups ( t 8 = 3.407 for SOX2, t 8 = 12.072 for KLF4, Student’s t test). E , the protein expression levels N-CADHERIN and VIMENTIN in shScramble- or shJARID2-transfected U251 cells. Values of protein levels were normalized to those of β-ACTIN levels. Values show the mean with S.D. (n = 5). ∗∗; p < 0.01 significant difference between the two groups ( t 8 = 9.389 for N-CADHERIN, t 8 = 6.029 for VIMENTIN, Student’s t test). F , decrease in the malignancy of U251 cells by downregulation of JARID2. shScramble or shJARID2 RNA expressing lentivirus transfected U251 cells were subcutaneously implanted in Balb/c-nude mice. Left graph shows Kaplan-Meier survival curves shScramble or shJARID2 RNA expressing lentivirus transfected U251 tumor-bearing mice (n = 7 for shScramble, n = 9 for shJARID2). Right graph shows the tumor volume of each individual mouse. ∗; p < 0.05 significant difference between the two groups (LogRank Holm-Sidak test).

Article Snippet: Membranes were reacted with antibodies against NAT10 (13365-1-AP, Proteintech, Wuhan, China, RRID:AB_2148944), p84 (THOC1, 10920-1-AP, Proteintech, RRID:AB_2202239), SOX2 (AF2018, R&D systems, RRID:AB_355110), KLF4 (#4038, Cell Signaling Technology, RRID:AB_2265207), OCT4 (#4038, Cell Signaling Technology), N-CADHERIN (AF6426, R&D systems, RRID:AB_10718850), VIMENTIN (MAB21052, R&D systems, RRID:AB_2832972), SUZ12 (#3737, Cell Signaling Technology, RRID:AB_2196850), JARID2 (#13594, Cell Signaling Technology, RRID:AB_2798269), THRAP3 (TRAP150, sc-133250, Santa Cruz Biotechnology, RRID:AB_2202901), MAD1L1 (MAD1, 18322-1-AP, Proteintech, RRID:AB_2139251) and β-ACTIN conjugated with horseradish peroxidase (sc-47778, Santa Cruz Biotechnology, RRID:AB_2202239).

Techniques: Transfection, shRNA, Staining, Expressing