Journal: Molecules

Article Title: Synthesis and Antiproliferative Activity against Cancer Cells of Indole-Aryl-Amide Derivatives

doi: 10.3390/molecules28010265

Figure Lengend Snippet: In vitro cytotoxic activity of new compounds tested on two normal and six cancer cell lines, expressed as IC 50 [μM] a .

Article Snippet: HT29 human colorectal adenocarcinoma cells, HeLa human cervix adenocarcinoma cells, MCF7 human breast adenocarcinoma cells, PC-3 human prostate adenocarcinoma cells, J6 Jurkat Clone E6-1 acute T cell leukemia, the healthy I407 human intestine cells, and mouse embryonic fibroblast 3T3 fibroblasts as controls cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA).

Techniques: In Vitro, Activity Assay

Journal: Molecules

Article Title: Synthesis and Antiproliferative Activity against Cancer Cells of Indole-Aryl-Amide Derivatives

doi: 10.3390/molecules28010265

Figure Lengend Snippet: Selectivity indices of compounds against cancer cell lines (IC 50 ) using 3T3 or I407 normal cells (IC 50 ) as reference.

Article Snippet: HT29 human colorectal adenocarcinoma cells, HeLa human cervix adenocarcinoma cells, MCF7 human breast adenocarcinoma cells, PC-3 human prostate adenocarcinoma cells, J6 Jurkat Clone E6-1 acute T cell leukemia, the healthy I407 human intestine cells, and mouse embryonic fibroblast 3T3 fibroblasts as controls cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA).

Techniques:

Journal: Frontiers in Immunology

Article Title: High-Mobility Group Box 1-Induced Complement Activation Causes Sterile Inflammation

doi: 10.3389/fimmu.2018.00705

Figure Lengend Snippet: High-mobility group box 1 (HMGB1)-mediated membrane attack complex (MAC) formation and its effect to cell signaling. (A,B) MEFs were cultured in DMEM containing 10% normal human serum (NHS) in the presence or absence of 1 µg/ml HMGB1 for 1 h at 37°C. Sublytic MAC proteins were stained using anti-C5b-9 antibody (Ab) (red) and observed by confocal microscopy. Blue: DAPI. Heat-inactivated (HI) NHS was used. Scale bar, 10 µm (A) . MEFs were incubated with 10% NHS in the presence of different concentrations of HMGB1, and then the mean relative intensity of fluorescence of 10 visual fields was calculated (B) . Error bars are mean ± SD. * P < 0.05 by Student’s paired t -test. (C) MEFs were cultured in DMEM containing 5% NHS in the presence of 1 µg/ml HMGB1 for 1 h at 37°C. To observe MAC formation, MEFs were fixed and mouse anti-C5b-9 Ab was used for immunofluorescent analysis. Cholera toxin B-FITC (CTB 0.5 µg/ml, Sigma) was utilized to observe the cell membrane lipid raft using confocal microscopy. Scale bar, 10 µm. (D) bEnd.3 cells were cultured in the presence of 5% NHS and/or 5 µg/ml HMGB1 and MAC formation was observed using Western blot analysis. (E,F) MAC formation. bEND.3 cells and LN215 cells were cocultured and incubated with DMEM containing 5% NHS in the presence or absence of 5 µg/ml HMGB1 for 16 h, and the alteration of tight junction (green line) and sublytic MAC deposition (red fluorescence, arrow) was observed by using confocal microscopy. Anti-ZO-1 Ab and anti-C5b-9 Abs were used for the study. Scale bar = 10 μm (E) . The mean relative intensity of fluorescence of six visual fields of zonal occluding (ZO) was calculated (F) . Error bars are mean ± SD. * P < 0.05 by Student’s paired t -test.

Article Snippet: Mouse embryonic fibroblasts (MEFs) (immortalized by the 3T3 protocol, purchased from HMGBiotech) were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 5–10% NHS 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM l -glutamine under 5% CO 2 in the presence or absence of 1 μg/ml HMGB1 for 1 h at 37°C.

Techniques: Membrane, Cell Culture, Staining, Confocal Microscopy, Incubation, Fluorescence, Western Blot