rpmi Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Lonza rpmi 1640
    Maraviroc inhibits crosstalk between cHL cells and both MSCs and monocytes. (A) Percentages of BM-MSCs and AT-MSCs that migrated (in 5 h) through a fibronectin-coated Boyden chamber towards conditioned medium (CM) from L-1236 or KM-H2 cells, in the presence of increasing concentrations of maraviroc (MSCs were treated for 1 h prior to migration). (B) Effect of a neutralizing anti-CCL5 antibody (5 μg/mL) in cHL-conditioned medium on MSC migration. Transmigrated cells were revealed using a computer-interfaced GeniusPlus microplate reader (Tecan). Results are the mean and SD of three replicate wells from three independent experiments. (C) BM-MSCs, AT-MSCs and HL-MSCs (100 cells/well; 24-well plates) were cultured in <t>RPMI-1640</t> medium containing 10% cHL CM. After 9 days, cells were fixed with methanol and stained with crystal violet. (D) BM-MSCs (500 cells/well; 96-well plates) were cultured in RPMI-1640 medium containing 20% cHL CM, with or without a neutralizing anti-FGF2 (1 μg/ml), anti-TGFβ1 (2 μg/mL) or anti-TNFα (0.5 μg/mL) antibody. After 9 days, growth was evaluated using the MTT assay. Results are the mean and SD of three replicate wells from three independent experiments. (E) BM-MSCs were cultured for 6 days with 20% CM from L-1236, L-428, KM-H2, HDLM-2, and L-540 cells. Then, the medium was changed with fresh medium, and 3 days later MSC CM was recovered and quantified for CCL5 by ELISA. All samples were tested in triplicate; conditioned media from three different experiments were evaluated. (F) BM-MSCs were cultured for 6 days with 20% CM (from KM-H2 or HDLM-2 cells) in the presence or absence of a neutralizing anti-TNFα antibody (0.5 μg/mL). Then, the medium was changed and after 3 days CCL5 was quantified by ELISA. All samples were run in triplicate; conditioned media from three different experiments were evaluated. (G) Clonogenic growth. L-1236 (10 3 /mL), HDLM-2 (5 × 10 2 /mL), L-540 (2.5 × 10 2 /mL) cells were cultured in methylcellulose-containing medium in the absence or presence of 5% E-BM-MSC CM and with increasing concentrations of maraviroc. After 14 days of incubation, plates were observed under phase contrast microscopy and aggregates with 40 cells or more were scored as colonies (8 replicate wells). Each experiment was done in triplicate; conditioned media from three different experiments were evaluated. (H) Percentage of CD14 + monocytes that migrated (in 1 h) through fibronectin-coated Boyden chambers towards medium (RPMI-1640 plus 10% FCS) or E-BM-MSC CM. Prior to migration towards E-BM-MSC CM, monocytes were pretreated with maraviroc (0.1-100 μM) for 1 h. Results are means and SD of transmigrated monocytes for three different experiments. AT-MSCs: Adipose Tissue; BM-MSCs: Bone Marrow; cHL: classical Hodgkin lymphoma; CM: conditioned medium; HL-MSCs: Hodgkin lymphoma; MSC: Mesenchymal stromal cells; E-BM-MSCs: tumor Educated.
    Rpmi 1640, supplied by Lonza, used in various techniques. Bioz Stars score: 99/100, based on 5862 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/Lonza
    Average 99 stars, based on 5862 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    ATCC rpmi 1640 medium
    Expression of macrophage-associated cytokines in THP-1 cells. Notes: ( A ) THP-1 cells are activated using PMA for 3 days and then treated with conditioned medium from different cells for 3 days. The mRNA is purified, and the expression of macrophage-associated cytokines (OPN, IL-10, CCL18, CD163, and CCL22) is assessed using RT-PCR (n = 2). The negative control of THP-1 is treated with <t>RPMI-1640</t> medium and 1% FBS. ( B ) Semiquantitative analysis of cDNA is performed, and β-actin is served as the loading control. The feature of M2 macrophages in THP-1 cells is increased after stimulation with conditioned medium from ampullary cancer cells. The experiments are repeated twice because of the scarcity of conditioned medium from primary culture cells. FHs 74 Int, a cell line from normal epithelial cells of the small intestine. AC01, primary culture cells from ampullary adenoma. AC02, primary culture cells from stage Ia ampullary cancer. AC03, primary culture cells from stage Ib ampullary cancer. AC04, primary culture cells from stage IIb ampullary cancer. TGBC-18-TKB, cell line from ampullary cancer. Abbreviations: CCL18, C-C motif ligand 18; CCL22, C-C motif ligand 22; CD163, cluster of differentiation 163; C.M, conditioned medium; FBS, fetal bovine serum; IL-10, interleukin-10; OPN, osteopontin; PMA, phorbol 12-myristate 13-acetate; RPMI, Roswell Park Memorial Institute; RT-PCR, reverse transcription-polymerase chain reaction.
    Rpmi 1640 Medium, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 2163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640 medium/product/ATCC
    Average 99 stars, based on 2163 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 medium - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher rpmi 1640 medium
    Block of fibronectin decreased the binding of M. hyopneumoniae to STEC. Adhesion rate: The number of bacteria recovered in the cells incubated with anti-fibronectin antibody/number of bacteria recovered in the cells incubated with <t>RPMI-1640</t> medium × 100%. Data are expressed as means ± SD of at least three experiments with samples performed in triplicate. The asterisk stands for statistically significant differences.
    Rpmi 1640 Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 100450 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640 medium/product/Thermo Fisher
    Average 99 stars, based on 100450 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 medium - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    96
    Thermo Fisher rpmi 1640
    Pyruvate impairs sensitivity to glutaminase inhibition by increasing TCA cycle anaplerosis. Increasing sodium pyruvate concentration in <t>RPMI</t> 1640 + 5% FBS increases CB-839 IC 50 (mean ± SEM, n = 3–7, one-way ANOVA) in a MDA-MB-231, b BT549 or c Hs578T cells during a 3 day assay. 24 h treatment with CB-839 decreased fumarate level in MDA-MB-231 cells in RPMI 1640 + 5% FBS but not RPMI 1640 + 5% FBS supplemented with 1 mM sodium pyruvate. Fumarate level was normalised to either initial cell number seeded ( d ) or protein amount at endpoint ( e ) (mean ± SEM, n = 3–4, two-way ANOVA)
    Rpmi 1640, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 96274 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/Thermo Fisher
    Average 96 stars, based on 96274 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    96/100 stars
      Buy from Supplier

    99
    Millipore rpmi 1640 medium
    Temsirolimus inhibited VEGF production by cancer cells and endothelial proliferation. ( A ) Tumor cells were incubated with or without HGF (50 ng/ml) in the presence of different concentrations of temsirolimus for 48 h. Then, supernatants were harvested and the number of tumor cells was counted. VEGF concentration in the supernatants was determined by ELISA. VEGF levels corrected by the tumor cell number are shown. ( B ) HMVECs were incubated in <t>RPMI-1640</t> medium with 10% FBS (control), RPMI-1640 medium with 10%-FBS plus VEGF, or HuMedia-MvG with different concentrations of temsirolimus for 72 h. Then, cell viability was determined using the MTT assay. Bars show SD. The data shown are 2 independent experiments with similar results.
    Rpmi 1640 Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 30869 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640 medium/product/Millipore
    Average 99 stars, based on 30869 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 medium - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Millipore rpmi 1640
    Activities of antioxidant enzymes in MDA-MB-23 and MCF-7 cells treated with 100 μg/ml of CE. Breast cancer cell lines, MCF-7 and MDA-MB-231, were seeded into 12-well plates containing RPMI 1640 and DMEM, respectively, and supplemented with 10% FBS at 5 × 10 6  cells/well and allowed to attach for 24 h. The cells were treated with 100 μg/ml of CE (IC 70  concentration determined from MTT assay) at varying time points (6, 9, 12, 24, and 48 h incubation). Activity of (A) catalase, (B) superoxide dismutase and (C) glutathione peroxidase was determined using commercial assay kits. Results are expressed as mean ± standard deviation. P
    Rpmi 1640, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 26338 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/Millipore
    Average 99 stars, based on 26338 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher rpmi medium
    Altered HIF1α-dependent effects of normoxia and hypoxia under different culture conditions. Western blot analysis was performed 24 h after the application of different treatments for HIF1α and carbonic anhydrase (CAIX), while β-actin served as a control. (Lane 1) Medium control; (lane 2) siRNA against HIF1α; (lane 3) 1% fetal bovine serum; (lane 4) 1% fetal bovine serum and siRNA against HIF1α; (lane 5) 5 mM glutamine; (lane 6) 5 mM glutamine and siRNA against HIF1α; (lane 7) 1% fetal bovine serum and 5 mM glutamine; and, (lane 8) 1% fetal bovine serum, 5 mM glutamine and siRNA against HIF1α under <t>normoxic</t> or hypoxic (*) conditions in MDA-MB-231 cells. The experiments were performed in <t>RPMI</t> 1640 in the presence of 11.1 mM glucose, but without glutamine. Deep sequencing analysis was performed for each of the corresponding RNA samples (lanes 7, 8, 7*, and 8*) from four independent experiments (Please see Supplemental Figure S3 and the results from the densitometric evaluation of the Western blots of four independent experiments).
    Rpmi Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 17348 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi medium/product/Thermo Fisher
    Average 99 stars, based on 17348 article reviews
    Price from $9.99 to $1999.99
    rpmi medium - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    GE Healthcare rpmi 1640 medium
    (A) LDH release from MCF-7 cells subjected to 2 μmol/L fluopsin C treatment. MCF-7 cells were treated with 1% Triton-X-100 (Sigma) in <t>RPMI-1640</t> medium for 10 min as the 100% LDH release control. b P
    Rpmi 1640 Medium, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 12494 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640 medium/product/GE Healthcare
    Average 99 stars, based on 12494 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 medium - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    GE Healthcare rpmi 1640
    Interaction of ghA, ghB, and ghC with monocyte/macrophages . PMBCs (1 × 10 6 ) were seeded on 13 mm coverslips and incubated in complete <t>RPMI</t> 1640 medium for 2 weeks at 37°C in 5% CO 2 incubator. Cells were treated with 10 μg of each globular head module and incubated with serum-free RPMI 1640 medium for 1 h at 37°C. After washing with PBS, cells were fixed with 4% PFA, permeabilzed with Triton X-100, and probed with anti-gC1qR polyclonal antibody and anti-MBP monoclonal antibody to reveal gC1qR and bound globular head modules, respectively. Cells were washed and treated with Alexa Fluor 488 conjugated secondary goat anti-rabbit antibody and Alexa Fluor 647 conjugated secondary donkey anti-mouse antibody and the nucleus was stained with Hoechst 33342. Cells were then examined under Leica fluorescence microscope with 40× magnification. In the merged images gC1qR is green; globular heads are red; and nucleus is blue. Arrows point to bound globular heads with colocalization of globular heads and gC1qR seen in orange in the merged images. Scale bars: 10 μm.
    Rpmi 1640, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 9823 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/GE Healthcare
    Average 99 stars, based on 9823 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    93
    Mediatech rpmi 1640
    Anti-tumor effect of G47Δ in NPC xenografts in mice. CNE-2 cells (1 × 10 6 ) and SUNE-1 cells (5 × 10 5 ) were suspended in 100 mL <t>RPMI-1640</t> complete medium with 25% Matrigel (BD Biosciences) and implanted subcutaneous into the left flanks of 4-week-old nude mice. When the maximal diameter of the subcutaneous tumors was approximately 5 mm, G47Δ (2 × 10 7 pfu /50 µL) or virus buffer (150 mmol/L NaCl and 20 mmol/L Tris at pH7.5) was injected into the subcutaneous tumors twice a week for 2 weeks. A, the subcutaneous CNE-2 tumor regressed by treatment with G47Δ on day 34 or appeared as large tumors after the mock treatment on day 20 post-injection of tumor cells. B, the subcutaneous SUNE-1 tumor regressed by treatment with G47Δ or appeared as large tumors after the mock treatment on day 37 post-injection of tumor cells. Comparison of mice with subcutaneous CNE-2 tumors (C) or subcutaneous SUNE-1 tumors (D) in the mock-treated group (upper) and the virus-treated group (lower) on day 20 or day 37 post-injection of tumor cells. Hematoxylin and eosin (HE) staining of CNE-2 (E), SUNE-1 (F), G47Δ-treated CNE-2 (G), and G47Δ-treated SUNE-1 (H) subcutaneous tumors (×200).
    Rpmi 1640, supplied by Mediatech, used in various techniques. Bioz Stars score: 93/100, based on 3945 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/Mediatech
    Average 93 stars, based on 3945 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    94
    Cellgro rpmi 1640
    Scanning electron photomicrographs of METs formed by bovine macrophages in response to M. haemolytica cells. Bovine macrophages (2.5 × 10 5 ) were incubated with 5 × 10 7 M. haemolytica cells or <t>RPMI</t> 1640 (negative control) for 5 min at 37°C.
    Rpmi 1640, supplied by Cellgro, used in various techniques. Bioz Stars score: 94/100, based on 2551 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/Cellgro
    Average 94 stars, based on 2551 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    94
    Corning Life Sciences rpmi 1640
    IMPDH forms filaments during ex vivo primary human T cell activation. (A) Representative images of T cells left untreated or stimulated by mitogens PHA, ConA, or anti-CD3/CD28 for 72 h, then fixed and stained for IMPDH (green) and T cell marker CD3 (red). White arrows: examples of IMPDH filaments. Yellow arrows: example of a ring-shaped filament. Images were captured using identical microscope settings for all treatment groups. (B) Quantification of the percentage of T cells that form filaments when untreated or treated with mitogens PHA, ConA, or anti-CD3/CD28. Cells were cultured in <t>RPMI</t> 1640 under four different conditions: 2 mM glutamine (Gln), 2 mM Gln + 1 h fresh medium, 16 mM Gln, or 16 mM Gln + 1 h fresh medium (represented by differently shaded bars). Different culture conditions were grouped together according to mitogenic treatment and compared to untreated cells (e.g., all PHA-treated conditions grouped vs. all untreated conditions grouped) and statistical significance displayed above each group. No significant differences were observed within these treatment groups due to culture conditions (e.g., PHA 2 mM Gln vs. PHA 16 mM Gln, no difference). Statistical test used: two-way ANOVA followed by Tukey's multiple comparisons test; **** p
    Rpmi 1640, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 94/100, based on 2500 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/Corning Life Sciences
    Average 94 stars, based on 2500 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    94
    Mediatech rpmi 1640 medium
    Compound 6-B345TTQ increases α4-dependent cell spreading in Jurkat T cells and CHO cells. Jurkat T cells expressing wild-type α4 ( A ) or α4(S988A) integrin-expressing CHO cells ( B ) were resuspended in <t>RPMI</t> 1640 medium or Dulbecco's
    Rpmi 1640 Medium, supplied by Mediatech, used in various techniques. Bioz Stars score: 94/100, based on 2810 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640 medium/product/Mediatech
    Average 94 stars, based on 2810 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 medium - by Bioz Stars, 2020-09
    94/100 stars
      Buy from Supplier

    99
    ATCC rpmi 1640
    Foam cell formation in hMDMs. Similar to oxLDL uptake studies, ether-linked AMs ( 3 ) displayed significantly higher efficacy than ester-linked AMs ( 5 ) at preventing the foam cell phenotype from developing, minimizing intracellular lipid accumulation. hMDMs were incubated with 50 µg/mL oxLDL and 10 −5 M AMs in <t>RPMI</t> 1640 for 24 h before staining with Oil Red O and Hoechst 33342.
    Rpmi 1640, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1528 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rpmi 1640/product/ATCC
    Average 99 stars, based on 1528 article reviews
    Price from $9.99 to $1999.99
    rpmi 1640 - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Maraviroc inhibits crosstalk between cHL cells and both MSCs and monocytes. (A) Percentages of BM-MSCs and AT-MSCs that migrated (in 5 h) through a fibronectin-coated Boyden chamber towards conditioned medium (CM) from L-1236 or KM-H2 cells, in the presence of increasing concentrations of maraviroc (MSCs were treated for 1 h prior to migration). (B) Effect of a neutralizing anti-CCL5 antibody (5 μg/mL) in cHL-conditioned medium on MSC migration. Transmigrated cells were revealed using a computer-interfaced GeniusPlus microplate reader (Tecan). Results are the mean and SD of three replicate wells from three independent experiments. (C) BM-MSCs, AT-MSCs and HL-MSCs (100 cells/well; 24-well plates) were cultured in RPMI-1640 medium containing 10% cHL CM. After 9 days, cells were fixed with methanol and stained with crystal violet. (D) BM-MSCs (500 cells/well; 96-well plates) were cultured in RPMI-1640 medium containing 20% cHL CM, with or without a neutralizing anti-FGF2 (1 μg/ml), anti-TGFβ1 (2 μg/mL) or anti-TNFα (0.5 μg/mL) antibody. After 9 days, growth was evaluated using the MTT assay. Results are the mean and SD of three replicate wells from three independent experiments. (E) BM-MSCs were cultured for 6 days with 20% CM from L-1236, L-428, KM-H2, HDLM-2, and L-540 cells. Then, the medium was changed with fresh medium, and 3 days later MSC CM was recovered and quantified for CCL5 by ELISA. All samples were tested in triplicate; conditioned media from three different experiments were evaluated. (F) BM-MSCs were cultured for 6 days with 20% CM (from KM-H2 or HDLM-2 cells) in the presence or absence of a neutralizing anti-TNFα antibody (0.5 μg/mL). Then, the medium was changed and after 3 days CCL5 was quantified by ELISA. All samples were run in triplicate; conditioned media from three different experiments were evaluated. (G) Clonogenic growth. L-1236 (10 3 /mL), HDLM-2 (5 × 10 2 /mL), L-540 (2.5 × 10 2 /mL) cells were cultured in methylcellulose-containing medium in the absence or presence of 5% E-BM-MSC CM and with increasing concentrations of maraviroc. After 14 days of incubation, plates were observed under phase contrast microscopy and aggregates with 40 cells or more were scored as colonies (8 replicate wells). Each experiment was done in triplicate; conditioned media from three different experiments were evaluated. (H) Percentage of CD14 + monocytes that migrated (in 1 h) through fibronectin-coated Boyden chambers towards medium (RPMI-1640 plus 10% FCS) or E-BM-MSC CM. Prior to migration towards E-BM-MSC CM, monocytes were pretreated with maraviroc (0.1-100 μM) for 1 h. Results are means and SD of transmigrated monocytes for three different experiments. AT-MSCs: Adipose Tissue; BM-MSCs: Bone Marrow; cHL: classical Hodgkin lymphoma; CM: conditioned medium; HL-MSCs: Hodgkin lymphoma; MSC: Mesenchymal stromal cells; E-BM-MSCs: tumor Educated.

    Journal: Haematologica

    Article Title: CCR5 antagonism by maraviroc inhibits Hodgkin lymphoma microenvironment interactions and xenograft growth

    doi: 10.3324/haematol.2018.196725

    Figure Lengend Snippet: Maraviroc inhibits crosstalk between cHL cells and both MSCs and monocytes. (A) Percentages of BM-MSCs and AT-MSCs that migrated (in 5 h) through a fibronectin-coated Boyden chamber towards conditioned medium (CM) from L-1236 or KM-H2 cells, in the presence of increasing concentrations of maraviroc (MSCs were treated for 1 h prior to migration). (B) Effect of a neutralizing anti-CCL5 antibody (5 μg/mL) in cHL-conditioned medium on MSC migration. Transmigrated cells were revealed using a computer-interfaced GeniusPlus microplate reader (Tecan). Results are the mean and SD of three replicate wells from three independent experiments. (C) BM-MSCs, AT-MSCs and HL-MSCs (100 cells/well; 24-well plates) were cultured in RPMI-1640 medium containing 10% cHL CM. After 9 days, cells were fixed with methanol and stained with crystal violet. (D) BM-MSCs (500 cells/well; 96-well plates) were cultured in RPMI-1640 medium containing 20% cHL CM, with or without a neutralizing anti-FGF2 (1 μg/ml), anti-TGFβ1 (2 μg/mL) or anti-TNFα (0.5 μg/mL) antibody. After 9 days, growth was evaluated using the MTT assay. Results are the mean and SD of three replicate wells from three independent experiments. (E) BM-MSCs were cultured for 6 days with 20% CM from L-1236, L-428, KM-H2, HDLM-2, and L-540 cells. Then, the medium was changed with fresh medium, and 3 days later MSC CM was recovered and quantified for CCL5 by ELISA. All samples were tested in triplicate; conditioned media from three different experiments were evaluated. (F) BM-MSCs were cultured for 6 days with 20% CM (from KM-H2 or HDLM-2 cells) in the presence or absence of a neutralizing anti-TNFα antibody (0.5 μg/mL). Then, the medium was changed and after 3 days CCL5 was quantified by ELISA. All samples were run in triplicate; conditioned media from three different experiments were evaluated. (G) Clonogenic growth. L-1236 (10 3 /mL), HDLM-2 (5 × 10 2 /mL), L-540 (2.5 × 10 2 /mL) cells were cultured in methylcellulose-containing medium in the absence or presence of 5% E-BM-MSC CM and with increasing concentrations of maraviroc. After 14 days of incubation, plates were observed under phase contrast microscopy and aggregates with 40 cells or more were scored as colonies (8 replicate wells). Each experiment was done in triplicate; conditioned media from three different experiments were evaluated. (H) Percentage of CD14 + monocytes that migrated (in 1 h) through fibronectin-coated Boyden chambers towards medium (RPMI-1640 plus 10% FCS) or E-BM-MSC CM. Prior to migration towards E-BM-MSC CM, monocytes were pretreated with maraviroc (0.1-100 μM) for 1 h. Results are means and SD of transmigrated monocytes for three different experiments. AT-MSCs: Adipose Tissue; BM-MSCs: Bone Marrow; cHL: classical Hodgkin lymphoma; CM: conditioned medium; HL-MSCs: Hodgkin lymphoma; MSC: Mesenchymal stromal cells; E-BM-MSCs: tumor Educated.

    Article Snippet: To prepare conditioned medium (CM) from cHL cell lines, cells were seeded at 2.0×105 /ml in RPMI-1640 plus 10% FCS, and the medium was collected after 72 h. Human bone marrow-derived and adipose tissue-derived mesenchymal stromal cells (MSCs) (BM-MSCs and AT-MSC, respectively) were purchased from Lonza (Verviers, Belgium). cHL-MSCs from frozen lymph nodes were generated as described in Online Supplementary Methods .

    Techniques: Migration, Cell Culture, Staining, MTT Assay, Enzyme-linked Immunosorbent Assay, Incubation, Microscopy

    Maraviroc slows tumor cell growth and inhibits heterospheroid formation. (A) cHL cells (2×10 5  cells/ml) were cultured with increasing concentrations of maraviroc for 72 h, and viable cells were counted by trypan blue dye exclusion. Results are means and SD of three replicate wells from three independent experiments. (B) Percentages of cHL cells in the various cell cycle phases after a 24 h treatment with maraviroc (100 μM). Results are means and SD of at least three experiments. (C) Representative image of 3D heterospheroids generated by plating HDLM-2 cells (stained green with CFSE), MSCs (red with fluorescent DiI), and monocytes (blue with DiD) under non-adherent conditions (poly-HEMA-coated wells). (D) L-1236 or HDLM-2 cells, HL-MSCs and monocytes were cultured in RPMI 1640 medium plus 1% FCS, alone or in combination under non-adherent conditions. After 4 days, conditioned medium was collected for CCL5 ELISAs; three different experiments were evaluated. (E) Heterospheroids generated by co-cultivation of cHL cells (L-1236, HDLM-2, or L-540 cells) with HL-MSCs and monocytes under non-adherent conditions in the presence or absence of maraviroc (100 μM) and photographed after 24 h using an inverted microscope (Eclipse TS/100, Nikon). (F) Heterospheroids (cHL + HL-MSCs + monocytes) were cultured with and without maraviroc. After 48 h, viable cells were evaluated using PrestoBlue Cell Viability Reagent. Relative fluorescence units (RFU). Values are means and SD of three experiments. cHL: classical Hodgkin lymphoma; (HL)-MSCs: Hodgkin lymphoma; MSC: mesenchymal stromal cells; 3D: three dimensional.

    Journal: Haematologica

    Article Title: CCR5 antagonism by maraviroc inhibits Hodgkin lymphoma microenvironment interactions and xenograft growth

    doi: 10.3324/haematol.2018.196725

    Figure Lengend Snippet: Maraviroc slows tumor cell growth and inhibits heterospheroid formation. (A) cHL cells (2×10 5 cells/ml) were cultured with increasing concentrations of maraviroc for 72 h, and viable cells were counted by trypan blue dye exclusion. Results are means and SD of three replicate wells from three independent experiments. (B) Percentages of cHL cells in the various cell cycle phases after a 24 h treatment with maraviroc (100 μM). Results are means and SD of at least three experiments. (C) Representative image of 3D heterospheroids generated by plating HDLM-2 cells (stained green with CFSE), MSCs (red with fluorescent DiI), and monocytes (blue with DiD) under non-adherent conditions (poly-HEMA-coated wells). (D) L-1236 or HDLM-2 cells, HL-MSCs and monocytes were cultured in RPMI 1640 medium plus 1% FCS, alone or in combination under non-adherent conditions. After 4 days, conditioned medium was collected for CCL5 ELISAs; three different experiments were evaluated. (E) Heterospheroids generated by co-cultivation of cHL cells (L-1236, HDLM-2, or L-540 cells) with HL-MSCs and monocytes under non-adherent conditions in the presence or absence of maraviroc (100 μM) and photographed after 24 h using an inverted microscope (Eclipse TS/100, Nikon). (F) Heterospheroids (cHL + HL-MSCs + monocytes) were cultured with and without maraviroc. After 48 h, viable cells were evaluated using PrestoBlue Cell Viability Reagent. Relative fluorescence units (RFU). Values are means and SD of three experiments. cHL: classical Hodgkin lymphoma; (HL)-MSCs: Hodgkin lymphoma; MSC: mesenchymal stromal cells; 3D: three dimensional.

    Article Snippet: To prepare conditioned medium (CM) from cHL cell lines, cells were seeded at 2.0×105 /ml in RPMI-1640 plus 10% FCS, and the medium was collected after 72 h. Human bone marrow-derived and adipose tissue-derived mesenchymal stromal cells (MSCs) (BM-MSCs and AT-MSC, respectively) were purchased from Lonza (Verviers, Belgium). cHL-MSCs from frozen lymph nodes were generated as described in Online Supplementary Methods .

    Techniques: Cell Culture, Generated, Staining, Inverted Microscopy, Fluorescence

    SCGB3A2 binding to LLC cells through HS of SDC1. ( A ) Schematic model of a human protein array for the isolation process of candidate genes shown as a Venn diagram. ( B ) Co-immunoprecipitation assay of SCGB3A2-FLAG and SDC1-Myc-His in COS-1 cells. IP and western blotting were sequentially carried out using anti-FLAG and anti-Myc antibody, respectively. ( C ) SCGB3A2 and SDC1 immunostaining in the airway epithelial cells of adult wild-type mouse lungs. Counterstained with Hematoxylin. Bar = 10 µm. ( D ) Immunofluorescent staining of SDC1 in LLC and B16F10 cells grown in 10%FBS-RPMI 1640 medium for 24 hr. DAPI was used for nuclear staining. White arrow indicates SDC1 cell surface expression in LLC. Red arrowhead points to the staining at cell-cell junction and white arrowheads point to focal SDC1 staining near the nucleus in B16F10 cells. Bar = 20 µm. ( E ) Flow cytometric analysis for SDC1 expression on cell surfaces of LLC and B16F10 cells using PE conjugated anti-SDC1 ectodomain specific antibody. ( F ) Flow cytometric analysis for SCGB3A2 binding to LLC cells using anti-SCGB3A2 antibody. GST tagged mouse SCGB3A2 (3 µg) was incubated with LLC cells at 4 ˚C for 30 min. Cells were stained with rabbit anti-mouse SCGB3A2 antibody, followed by staining with PE-anti-rabbit IgG antibody at 4 ˚C for 30 min. ( G ) SCGB3A2 binding assay on LLC-sh-Control or sh-SDC1 cells. GST tagged mouse SCGB3A2 (1 µg) was incubated with each cell type at 4 ˚C for 30 min, followed by staining with Alexa 488 anti-rabbit IgG antibody at 4 ˚C for 30 min. ( H ) SCGB3A2 binding assay on LLC cells. Cells were co-incubated with or without GST tagged mouse SCGB3A2 (1 µg) or SCGB3A2 + heparin. Cells were stained with PE-anti-rabbit IgG antibody at 4 ˚C for 30 min. Data except A are the representative from more than three independent experiments.

    Journal: eLife

    Article Title: A novel pathway of LPS uptake through syndecan-1 leading to pyroptotic cell death

    doi: 10.7554/eLife.37854

    Figure Lengend Snippet: SCGB3A2 binding to LLC cells through HS of SDC1. ( A ) Schematic model of a human protein array for the isolation process of candidate genes shown as a Venn diagram. ( B ) Co-immunoprecipitation assay of SCGB3A2-FLAG and SDC1-Myc-His in COS-1 cells. IP and western blotting were sequentially carried out using anti-FLAG and anti-Myc antibody, respectively. ( C ) SCGB3A2 and SDC1 immunostaining in the airway epithelial cells of adult wild-type mouse lungs. Counterstained with Hematoxylin. Bar = 10 µm. ( D ) Immunofluorescent staining of SDC1 in LLC and B16F10 cells grown in 10%FBS-RPMI 1640 medium for 24 hr. DAPI was used for nuclear staining. White arrow indicates SDC1 cell surface expression in LLC. Red arrowhead points to the staining at cell-cell junction and white arrowheads point to focal SDC1 staining near the nucleus in B16F10 cells. Bar = 20 µm. ( E ) Flow cytometric analysis for SDC1 expression on cell surfaces of LLC and B16F10 cells using PE conjugated anti-SDC1 ectodomain specific antibody. ( F ) Flow cytometric analysis for SCGB3A2 binding to LLC cells using anti-SCGB3A2 antibody. GST tagged mouse SCGB3A2 (3 µg) was incubated with LLC cells at 4 ˚C for 30 min. Cells were stained with rabbit anti-mouse SCGB3A2 antibody, followed by staining with PE-anti-rabbit IgG antibody at 4 ˚C for 30 min. ( G ) SCGB3A2 binding assay on LLC-sh-Control or sh-SDC1 cells. GST tagged mouse SCGB3A2 (1 µg) was incubated with each cell type at 4 ˚C for 30 min, followed by staining with Alexa 488 anti-rabbit IgG antibody at 4 ˚C for 30 min. ( H ) SCGB3A2 binding assay on LLC cells. Cells were co-incubated with or without GST tagged mouse SCGB3A2 (1 µg) or SCGB3A2 + heparin. Cells were stained with PE-anti-rabbit IgG antibody at 4 ˚C for 30 min. Data except A are the representative from more than three independent experiments.

    Article Snippet: LLC and B16F10 cells were cultured in RPMI 1640 Medium (LONZA) with heat-inactivated fetal bovine serum (FBS), supplemented with penicillin/streptomycin (1:100) at 37 ˚C, 5% CO2 .

    Techniques: Binding Assay, Protein Array, Isolation, Co-Immunoprecipitation Assay, Western Blot, Immunostaining, Staining, Expressing, Flow Cytometry, Incubation

    SCGB3A2-LPS promotion of pyroptotic cell death. ( A ) Representative phase contrast image of LLC cells incubated with LPS (O111:B4, 1 µg/ml) and SCGB3A2 (1 µg/ml) in 1% FBS-RPMI 1640 for 72 hr. Arrows indicate cells undergoing pyroptosis (swollen cells). Bar = 10 µm. Image is the representative of three independent experiments ( B ) (10 ng/ml) in 2%FBS-RPMI for 72 hr culture. Data are the representative from more than three independent experiments, each in triplicate. Averages ± SD are shown. (**p

    Journal: eLife

    Article Title: A novel pathway of LPS uptake through syndecan-1 leading to pyroptotic cell death

    doi: 10.7554/eLife.37854

    Figure Lengend Snippet: SCGB3A2-LPS promotion of pyroptotic cell death. ( A ) Representative phase contrast image of LLC cells incubated with LPS (O111:B4, 1 µg/ml) and SCGB3A2 (1 µg/ml) in 1% FBS-RPMI 1640 for 72 hr. Arrows indicate cells undergoing pyroptosis (swollen cells). Bar = 10 µm. Image is the representative of three independent experiments ( B ) (10 ng/ml) in 2%FBS-RPMI for 72 hr culture. Data are the representative from more than three independent experiments, each in triplicate. Averages ± SD are shown. (**p

    Article Snippet: LLC and B16F10 cells were cultured in RPMI 1640 Medium (LONZA) with heat-inactivated fetal bovine serum (FBS), supplemented with penicillin/streptomycin (1:100) at 37 ˚C, 5% CO2 .

    Techniques: Incubation

    Evaluation for the requirement of SDC1 and other genes for SCGB3A2-LPS effect. ( A ) CCK8 assay using LLC-sh-Control and LLC-sh-SDC1 for 72 hr in 1% FBS-RPMI 1640 medium. C; control, S; human SCGB3A2 (200 ng/ml), L; LPS (O111:B4, 1 pg/ml). Averages ± SD from more than three independent experiments, each in triplicate are shown. *: p

    Journal: eLife

    Article Title: A novel pathway of LPS uptake through syndecan-1 leading to pyroptotic cell death

    doi: 10.7554/eLife.37854

    Figure Lengend Snippet: Evaluation for the requirement of SDC1 and other genes for SCGB3A2-LPS effect. ( A ) CCK8 assay using LLC-sh-Control and LLC-sh-SDC1 for 72 hr in 1% FBS-RPMI 1640 medium. C; control, S; human SCGB3A2 (200 ng/ml), L; LPS (O111:B4, 1 pg/ml). Averages ± SD from more than three independent experiments, each in triplicate are shown. *: p

    Article Snippet: LLC and B16F10 cells were cultured in RPMI 1640 Medium (LONZA) with heat-inactivated fetal bovine serum (FBS), supplemented with penicillin/streptomycin (1:100) at 37 ˚C, 5% CO2 .

    Techniques: CCK-8 Assay

    Expression of macrophage-associated cytokines in THP-1 cells. Notes: ( A ) THP-1 cells are activated using PMA for 3 days and then treated with conditioned medium from different cells for 3 days. The mRNA is purified, and the expression of macrophage-associated cytokines (OPN, IL-10, CCL18, CD163, and CCL22) is assessed using RT-PCR (n = 2). The negative control of THP-1 is treated with RPMI-1640 medium and 1% FBS. ( B ) Semiquantitative analysis of cDNA is performed, and β-actin is served as the loading control. The feature of M2 macrophages in THP-1 cells is increased after stimulation with conditioned medium from ampullary cancer cells. The experiments are repeated twice because of the scarcity of conditioned medium from primary culture cells. FHs 74 Int, a cell line from normal epithelial cells of the small intestine. AC01, primary culture cells from ampullary adenoma. AC02, primary culture cells from stage Ia ampullary cancer. AC03, primary culture cells from stage Ib ampullary cancer. AC04, primary culture cells from stage IIb ampullary cancer. TGBC-18-TKB, cell line from ampullary cancer. Abbreviations: CCL18, C-C motif ligand 18; CCL22, C-C motif ligand 22; CD163, cluster of differentiation 163; C.M, conditioned medium; FBS, fetal bovine serum; IL-10, interleukin-10; OPN, osteopontin; PMA, phorbol 12-myristate 13-acetate; RPMI, Roswell Park Memorial Institute; RT-PCR, reverse transcription-polymerase chain reaction.

    Journal: OncoTargets and therapy

    Article Title: Cancer-Derived Transforming Growth Factor-β Modulates Tumor-Associated Macrophages in Ampullary Cancer

    doi: 10.2147/OTT.S246714

    Figure Lengend Snippet: Expression of macrophage-associated cytokines in THP-1 cells. Notes: ( A ) THP-1 cells are activated using PMA for 3 days and then treated with conditioned medium from different cells for 3 days. The mRNA is purified, and the expression of macrophage-associated cytokines (OPN, IL-10, CCL18, CD163, and CCL22) is assessed using RT-PCR (n = 2). The negative control of THP-1 is treated with RPMI-1640 medium and 1% FBS. ( B ) Semiquantitative analysis of cDNA is performed, and β-actin is served as the loading control. The feature of M2 macrophages in THP-1 cells is increased after stimulation with conditioned medium from ampullary cancer cells. The experiments are repeated twice because of the scarcity of conditioned medium from primary culture cells. FHs 74 Int, a cell line from normal epithelial cells of the small intestine. AC01, primary culture cells from ampullary adenoma. AC02, primary culture cells from stage Ia ampullary cancer. AC03, primary culture cells from stage Ib ampullary cancer. AC04, primary culture cells from stage IIb ampullary cancer. TGBC-18-TKB, cell line from ampullary cancer. Abbreviations: CCL18, C-C motif ligand 18; CCL22, C-C motif ligand 22; CD163, cluster of differentiation 163; C.M, conditioned medium; FBS, fetal bovine serum; IL-10, interleukin-10; OPN, osteopontin; PMA, phorbol 12-myristate 13-acetate; RPMI, Roswell Park Memorial Institute; RT-PCR, reverse transcription-polymerase chain reaction.

    Article Snippet: The THP-1 monocytes were maintained in a RPMI-1640 medium with 10% FBS.

    Techniques: Expressing, Purification, Reverse Transcription Polymerase Chain Reaction, Negative Control

    Block of fibronectin decreased the binding of M. hyopneumoniae to STEC. Adhesion rate: The number of bacteria recovered in the cells incubated with anti-fibronectin antibody/number of bacteria recovered in the cells incubated with RPMI-1640 medium × 100%. Data are expressed as means ± SD of at least three experiments with samples performed in triplicate. The asterisk stands for statistically significant differences.

    Journal: Frontiers in Microbiology

    Article Title: Elongation Factor Thermo Unstable (EF-Tu) Moonlights as an Adhesin on the Surface of Mycoplasma hyopneumoniae by Binding to Fibronectin

    doi: 10.3389/fmicb.2018.00974

    Figure Lengend Snippet: Block of fibronectin decreased the binding of M. hyopneumoniae to STEC. Adhesion rate: The number of bacteria recovered in the cells incubated with anti-fibronectin antibody/number of bacteria recovered in the cells incubated with RPMI-1640 medium × 100%. Data are expressed as means ± SD of at least three experiments with samples performed in triplicate. The asterisk stands for statistically significant differences.

    Article Snippet: M. hyopneumoniae strain 168 was cultured in RPMI-1640 medium with 2% (v/v) FBS in 24-well plates for use as the control.

    Techniques: Blocking Assay, Binding Assay, Incubation

    Pyruvate impairs sensitivity to glutaminase inhibition by increasing TCA cycle anaplerosis. Increasing sodium pyruvate concentration in RPMI 1640 + 5% FBS increases CB-839 IC 50 (mean ± SEM, n = 3–7, one-way ANOVA) in a MDA-MB-231, b BT549 or c Hs578T cells during a 3 day assay. 24 h treatment with CB-839 decreased fumarate level in MDA-MB-231 cells in RPMI 1640 + 5% FBS but not RPMI 1640 + 5% FBS supplemented with 1 mM sodium pyruvate. Fumarate level was normalised to either initial cell number seeded ( d ) or protein amount at endpoint ( e ) (mean ± SEM, n = 3–4, two-way ANOVA)

    Journal: BMC Cancer

    Article Title: Pyruvate anaplerosis is a mechanism of resistance to pharmacological glutaminase inhibition in triple-receptor negative breast cancer

    doi: 10.1186/s12885-020-06885-3

    Figure Lengend Snippet: Pyruvate impairs sensitivity to glutaminase inhibition by increasing TCA cycle anaplerosis. Increasing sodium pyruvate concentration in RPMI 1640 + 5% FBS increases CB-839 IC 50 (mean ± SEM, n = 3–7, one-way ANOVA) in a MDA-MB-231, b BT549 or c Hs578T cells during a 3 day assay. 24 h treatment with CB-839 decreased fumarate level in MDA-MB-231 cells in RPMI 1640 + 5% FBS but not RPMI 1640 + 5% FBS supplemented with 1 mM sodium pyruvate. Fumarate level was normalised to either initial cell number seeded ( d ) or protein amount at endpoint ( e ) (mean ± SEM, n = 3–4, two-way ANOVA)

    Article Snippet: On day 1, 50 μL of fresh RPMI 1640 + 5% FBS was added to the plates.

    Techniques: Inhibition, Concentration Assay, Multiple Displacement Amplification

    Pyruvate secreted by TNBC cell lines reduces the potency of CB-839. a Pharmacological MCT1 inhibition using 1 μM AZD3965 reduced the secretion of pyruvate by Hs578T and SUM159PT (SUM159) but not MDA-MB-231-luc-D3H2LN (D3H2LN) cells. Pyruvate concentration in the conditioned or unconditioned RPMI 1640 + 5% FBS (RPMI) culture medium was quantified after 48 h incubation (mean ± SEM, n = 3, t test). b The pyruvate concentration in the conditioned culture medium from a correlates with resistance of recipient MDA-MB-231 cells to CB-839-treatment at 10 nM, 100 nM, 1 μM or 10 μM over 3 days exposure. For each of the TNBC cell lines studied the AZD3965-treated samples of conditioned culture medium demonstrated a decrease in relative thymidine incorporation in recipient MDA-MB-231 cells (mean ± SD, n = 2). c IC 50 analysis also demonstrates a correlation between CB-839 sensitivity and pyruvate concentration in the conditioned culture medium from a (mean ± SD, n = 2). Correlations were computed by Pearson r correlation coefficient analysis

    Journal: BMC Cancer

    Article Title: Pyruvate anaplerosis is a mechanism of resistance to pharmacological glutaminase inhibition in triple-receptor negative breast cancer

    doi: 10.1186/s12885-020-06885-3

    Figure Lengend Snippet: Pyruvate secreted by TNBC cell lines reduces the potency of CB-839. a Pharmacological MCT1 inhibition using 1 μM AZD3965 reduced the secretion of pyruvate by Hs578T and SUM159PT (SUM159) but not MDA-MB-231-luc-D3H2LN (D3H2LN) cells. Pyruvate concentration in the conditioned or unconditioned RPMI 1640 + 5% FBS (RPMI) culture medium was quantified after 48 h incubation (mean ± SEM, n = 3, t test). b The pyruvate concentration in the conditioned culture medium from a correlates with resistance of recipient MDA-MB-231 cells to CB-839-treatment at 10 nM, 100 nM, 1 μM or 10 μM over 3 days exposure. For each of the TNBC cell lines studied the AZD3965-treated samples of conditioned culture medium demonstrated a decrease in relative thymidine incorporation in recipient MDA-MB-231 cells (mean ± SD, n = 2). c IC 50 analysis also demonstrates a correlation between CB-839 sensitivity and pyruvate concentration in the conditioned culture medium from a (mean ± SD, n = 2). Correlations were computed by Pearson r correlation coefficient analysis

    Article Snippet: On day 1, 50 μL of fresh RPMI 1640 + 5% FBS was added to the plates.

    Techniques: Inhibition, Multiple Displacement Amplification, Concentration Assay, Incubation

    Breast cancer cell lines display differences in sensitivity to pharmacological glutaminase inhibition depending on culture medium composition. a MDA-MB-231 cells were more sensitive to 3 days CB-839 exposure when assayed in RPMI 1640 + 5% FBS (RPMI 1640) compared with αMEM + 5% FBS (αMEM) or DMEM + 5% FBS (DMEM) (mean ± SEM, n = 2–4, one-way ANOVA). CB-839 ( b ) or BPTES ( c ) display more potent IC 50 values when assayed in RPMI 1640 + 5% FBS compared with αMEM + 5% FBS in many breast cancer cell lines (mean ± SEM, n = 2–4, unpaired t test)

    Journal: BMC Cancer

    Article Title: Pyruvate anaplerosis is a mechanism of resistance to pharmacological glutaminase inhibition in triple-receptor negative breast cancer

    doi: 10.1186/s12885-020-06885-3

    Figure Lengend Snippet: Breast cancer cell lines display differences in sensitivity to pharmacological glutaminase inhibition depending on culture medium composition. a MDA-MB-231 cells were more sensitive to 3 days CB-839 exposure when assayed in RPMI 1640 + 5% FBS (RPMI 1640) compared with αMEM + 5% FBS (αMEM) or DMEM + 5% FBS (DMEM) (mean ± SEM, n = 2–4, one-way ANOVA). CB-839 ( b ) or BPTES ( c ) display more potent IC 50 values when assayed in RPMI 1640 + 5% FBS compared with αMEM + 5% FBS in many breast cancer cell lines (mean ± SEM, n = 2–4, unpaired t test)

    Article Snippet: On day 1, 50 μL of fresh RPMI 1640 + 5% FBS was added to the plates.

    Techniques: Inhibition, Multiple Displacement Amplification

    Lactoferrin enhancement of the maturation of peripheral blood dendritic cells (PBDCs) (A) PBMCs were cultured for 40 h in RPMI 1640 containing 5% FCS with or without TLF (100 μg/ml) or LPS (200 ng/ml). The cells were subsequently stained to analyze by flow cytometry CD80, CD83, and CD86 expression by gating on CD1c + PBDCs subset. Data shown are the results of one experiment representative of four. B) Purified CD1c + PBDCs were cultured (10 5 /well) in RPMI-5%FCS for 48 h in the presence of TLF (10 μg/ml) or LPS (500 ng/ml). The supernatants were collected for the measurement of cytokine production. The results of one experiment representative of two are shown.

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Lactoferrin acts as an alarmin to promote the recruitment and activation of antigen-presenting cells and antigen-specific immune responses

    doi:

    Figure Lengend Snippet: Lactoferrin enhancement of the maturation of peripheral blood dendritic cells (PBDCs) (A) PBMCs were cultured for 40 h in RPMI 1640 containing 5% FCS with or without TLF (100 μg/ml) or LPS (200 ng/ml). The cells were subsequently stained to analyze by flow cytometry CD80, CD83, and CD86 expression by gating on CD1c + PBDCs subset. Data shown are the results of one experiment representative of four. B) Purified CD1c + PBDCs were cultured (10 5 /well) in RPMI-5%FCS for 48 h in the presence of TLF (10 μg/ml) or LPS (500 ng/ml). The supernatants were collected for the measurement of cytokine production. The results of one experiment representative of two are shown.

    Article Snippet: Briefly, cells were resuspended in RPMI containing 10% FCS (GIBCO-Invitrogen, Carlsbad, CA) at 1×106 cells/ml in the presence of IL-4 and GM-CSF (Pepro Tech, both at 50 ng/ml) every two days.

    Techniques: Cell Culture, Staining, Flow Cytometry, Cytometry, Expressing, Purification

    COTL1 is required for lamellipodial protrusion at the immune synapse. ( A ) Cartoon depicting the suppression and suppression/re-expression plasmids utilized in these studies. ( B, C ) Immunoblot analysis showing that endogenous COTL1 protein was suppressed and fluorescent-tagged COTL1 WT and COTL1 ABM were reconstituted, respectively. ( D ) Jurkat T cells were transfected with the GFP-expressing suppression/re-expression plasmids shown in ( A ). 72 h post transfection they were conjugated with CMAC-stained Raji B cells loaded or unloaded with SEE. Cell conjugates were stained for F-actin and imaged using fluorescence microscopy. Representative images were shown ( D , top panels) and F-actin accumulation at the synapse was scored and quantification was displayed ( D , lower graph). Over 100 conjugates were scored in 3 independent experiments. Error bars represent SEM. ( E ) Jurkat T cells stably expressing GFP-actin were transfected with mCherry suppression/re-expression plasmids shown in ( A ) and cultured for 3 d. For live cell imaging, RPMI 1640 media containing CMAC-stained SEE-loaded Raji B cells were added to Poly-L-lysine-coated coverglass chamber and incubated for 10 min. Afterwards, Jurkat T cells were dropped above the chamber and cherry positive Jurkat T cells were monitored by confocal microscopy every 20 s for 30 min following Jurkat T-Raji B cell contact for GFP-actin enriched lamellipodial protrusion. Image sequences captured every 100 s were shown. See associated Movies S1 - S4 .

    Journal: PLoS ONE

    Article Title: Coactosin-Like 1 Antagonizes Cofilin to Promote Lamellipodial Protrusion at the Immune Synapse

    doi: 10.1371/journal.pone.0085090

    Figure Lengend Snippet: COTL1 is required for lamellipodial protrusion at the immune synapse. ( A ) Cartoon depicting the suppression and suppression/re-expression plasmids utilized in these studies. ( B, C ) Immunoblot analysis showing that endogenous COTL1 protein was suppressed and fluorescent-tagged COTL1 WT and COTL1 ABM were reconstituted, respectively. ( D ) Jurkat T cells were transfected with the GFP-expressing suppression/re-expression plasmids shown in ( A ). 72 h post transfection they were conjugated with CMAC-stained Raji B cells loaded or unloaded with SEE. Cell conjugates were stained for F-actin and imaged using fluorescence microscopy. Representative images were shown ( D , top panels) and F-actin accumulation at the synapse was scored and quantification was displayed ( D , lower graph). Over 100 conjugates were scored in 3 independent experiments. Error bars represent SEM. ( E ) Jurkat T cells stably expressing GFP-actin were transfected with mCherry suppression/re-expression plasmids shown in ( A ) and cultured for 3 d. For live cell imaging, RPMI 1640 media containing CMAC-stained SEE-loaded Raji B cells were added to Poly-L-lysine-coated coverglass chamber and incubated for 10 min. Afterwards, Jurkat T cells were dropped above the chamber and cherry positive Jurkat T cells were monitored by confocal microscopy every 20 s for 30 min following Jurkat T-Raji B cell contact for GFP-actin enriched lamellipodial protrusion. Image sequences captured every 100 s were shown. See associated Movies S1 - S4 .

    Article Snippet: Cells were resuspended in 10 ml of RPMI 1640 media supplemented with 5% FCS (Invitrogen).

    Techniques: Expressing, Transfection, Staining, Fluorescence, Microscopy, Stable Transfection, Cell Culture, Live Cell Imaging, Incubation, Confocal Microscopy

    Observation of C. albicans yeast and hyphal forms, and colony numbers after inoculation. C. albicans yeast and hyphal forms were taken pictures in microscope after incubation of them in YPD and RPMI1640 with 5% FBS ( A ). The colonization was observed on the YPD agar and BHI agar plates poured swab samples from oral cavity of NOD/SCID. e2f1 −/− mice fed 1% sucrose water at 60 min after the inoculation ( B ). White and black arrow indicated C. albicans and indigenous microorganism colony respectively. These colonies were taken pictures in the stereoscopic microscope. Data were representative in three independent assays. Samples were swabbed from the oral cavities of four 4-month-old female NOD/SCID. e2f1 +/+ and NOD/SCID. e2f1 −/− mice at 30, 60, and 90 minutes after inoculation using the yeast form. Samples swabbed in mice fed water ( C ) or 1% sucrose water ( D ) overnight before inoculation were poured on YPD agar. Samples swabbed in mice fed water ( E ) or 1% water ( F ) before inoculation were poured on BHI agar. CFU data were obtained from three independent experiments with four mice from each strain. Values are expressed as the means ± standard deviations (SDs) of the data. Asterisks show significant differences (* P

    Journal: BMC Oral Health

    Article Title: Effects of salivary protein flow and indigenous microorganisms on initial colonization of Candida albicans in an in vivo model

    doi: 10.1186/1472-6831-12-36

    Figure Lengend Snippet: Observation of C. albicans yeast and hyphal forms, and colony numbers after inoculation. C. albicans yeast and hyphal forms were taken pictures in microscope after incubation of them in YPD and RPMI1640 with 5% FBS ( A ). The colonization was observed on the YPD agar and BHI agar plates poured swab samples from oral cavity of NOD/SCID. e2f1 −/− mice fed 1% sucrose water at 60 min after the inoculation ( B ). White and black arrow indicated C. albicans and indigenous microorganism colony respectively. These colonies were taken pictures in the stereoscopic microscope. Data were representative in three independent assays. Samples were swabbed from the oral cavities of four 4-month-old female NOD/SCID. e2f1 +/+ and NOD/SCID. e2f1 −/− mice at 30, 60, and 90 minutes after inoculation using the yeast form. Samples swabbed in mice fed water ( C ) or 1% sucrose water ( D ) overnight before inoculation were poured on YPD agar. Samples swabbed in mice fed water ( E ) or 1% water ( F ) before inoculation were poured on BHI agar. CFU data were obtained from three independent experiments with four mice from each strain. Values are expressed as the means ± standard deviations (SDs) of the data. Asterisks show significant differences (* P

    Article Snippet: The yeast form of C. albicans was predominant in culture with YPD overnight; whereas the mycelial form occurred in RPMI1640 (Gibco-Invitrogen, Grand Island, NY) with 5% FBS in overnight culture (Figure A).

    Techniques: Microscopy, Incubation, Mouse Assay

    Growth curves of different C. tropicalis isolates with different fluconazole susceptibilities. Six isolates, ATCC13803 (rectangles), NHUE40 (diamonds), NHUE42 (triangles), NHUE48 (circles), YM990579 (crosses), and YM990649 (stars) were grown in the RPMI medium 1640 (Gibco BRL31800-022) in the absence (dot lines) or in the presence of 64 mg/L (solid lines) fluconazole.

    Journal: PLoS ONE

    Article Title: Comparison of Human and Soil Candida tropicalis Isolates with Reduced Susceptibility to Fluconazole

    doi: 10.1371/journal.pone.0034609

    Figure Lengend Snippet: Growth curves of different C. tropicalis isolates with different fluconazole susceptibilities. Six isolates, ATCC13803 (rectangles), NHUE40 (diamonds), NHUE42 (triangles), NHUE48 (circles), YM990579 (crosses), and YM990649 (stars) were grown in the RPMI medium 1640 (Gibco BRL31800-022) in the absence (dot lines) or in the presence of 64 mg/L (solid lines) fluconazole.

    Article Snippet: RPMI medium 1640 (31800-022, Gibco BRL) was used for the dilution and growth of the yeast culture.

    Techniques:

    Construction of S. oralis spxB deletion mutant. (A) Black arrow indicates the gene encoding pyruvate oxidase (SMSK23_0092 spxB). A targeted deletion mutant lacking this region was constructed by allelic exchange using the temperature-sensitive shuttle vector pSET4s. (B) S. oralis ATCC35037 wild-type (WT), spxB-deletion mutant (KO), or reverse mutant (Rev) was cultured in BHI broth or 5% RPMI1640 medium at 37°C for 18 h in a 5% CO 2 atmosphere. Concentrations of H 2 O 2 in culture supernatants were quantitatively determined using a hydrogen peroxide colorimetric detection kit. Data are shown as the mean ± SD of triplicate samples. *p

    Journal: PLoS ONE

    Article Title: Hydrogen Peroxide Produced by Oral Streptococci Induces Macrophage Cell Death

    doi: 10.1371/journal.pone.0062563

    Figure Lengend Snippet: Construction of S. oralis spxB deletion mutant. (A) Black arrow indicates the gene encoding pyruvate oxidase (SMSK23_0092 spxB). A targeted deletion mutant lacking this region was constructed by allelic exchange using the temperature-sensitive shuttle vector pSET4s. (B) S. oralis ATCC35037 wild-type (WT), spxB-deletion mutant (KO), or reverse mutant (Rev) was cultured in BHI broth or 5% RPMI1640 medium at 37°C for 18 h in a 5% CO 2 atmosphere. Concentrations of H 2 O 2 in culture supernatants were quantitatively determined using a hydrogen peroxide colorimetric detection kit. Data are shown as the mean ± SD of triplicate samples. *p

    Article Snippet: Cell culture The human monocyte cell line THP-1 cells were purchased from RIKEN Bioresource Center and cultured in RPMI1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 5% fetal bovine serum (FBS) (Invitrogen) (5% FBS RPMI1640), penicillin (100 U/ml), and streptomycin (100 µg/ml) at 37°C in a 5% CO2 atmosphere.

    Techniques: Mutagenesis, Construct, Plasmid Preparation, Cell Culture

    STAMP2 loss inhibits androgen-insensitive PCa cell growth in vitro and in vivo A 22Rv1 cells were cultured in RPMI 1640 medium containing 10% CT-FBS and treated with or without 1 nM R1881 for 24 h and were then transfected with the indicated siRNAs. The cells were then cultured for 10 days. The colonies formed were stained and photographed. B Quantification of data from (A). Student's t -test was performed to analyze the statistical significance, n = 3. * P = 0.0003; ** P

    Journal: EMBO Molecular Medicine

    Article Title: STAMP2 increases oxidative stress and is critical for prostate cancer

    doi: 10.15252/emmm.201404181

    Figure Lengend Snippet: STAMP2 loss inhibits androgen-insensitive PCa cell growth in vitro and in vivo A 22Rv1 cells were cultured in RPMI 1640 medium containing 10% CT-FBS and treated with or without 1 nM R1881 for 24 h and were then transfected with the indicated siRNAs. The cells were then cultured for 10 days. The colonies formed were stained and photographed. B Quantification of data from (A). Student's t -test was performed to analyze the statistical significance, n = 3. * P = 0.0003; ** P

    Article Snippet: LNCaP cells and VCaP cells (ATCC, LGC standard) were routinely maintained in RPMI 1640 culture medium supplemented with 10% fetal bovine serum (FBS), 5 mg/ml penicillin/streptomycin, and 2 mM l -glutamine (Invitrogen) at 37°C in a humidified atmosphere with 5% CO2 .

    Techniques: In Vitro, In Vivo, Cell Culture, Transfection, Staining

    STAMP2 promotes PCa growth in vitro and in vivo A LNCaP cells were transfected with either control siRNA or two independent STAMP2 siRNAs, and membrane fractions of the cells were prepared followed by Western blotting analysis. STAMP1 is used as a loading control. B LNCaP cells were cultured in RPMI 1640 medium containing 10% CT-FBS and treated with or without 1 nM R1881 for 24 h before being transfected with the indicated siRNAs. The cells were then cultured for the indicated times, and cell growth was measured by Cell Counting Kit-8. n = 3, * P

    Journal: EMBO Molecular Medicine

    Article Title: STAMP2 increases oxidative stress and is critical for prostate cancer

    doi: 10.15252/emmm.201404181

    Figure Lengend Snippet: STAMP2 promotes PCa growth in vitro and in vivo A LNCaP cells were transfected with either control siRNA or two independent STAMP2 siRNAs, and membrane fractions of the cells were prepared followed by Western blotting analysis. STAMP1 is used as a loading control. B LNCaP cells were cultured in RPMI 1640 medium containing 10% CT-FBS and treated with or without 1 nM R1881 for 24 h before being transfected with the indicated siRNAs. The cells were then cultured for the indicated times, and cell growth was measured by Cell Counting Kit-8. n = 3, * P

    Article Snippet: LNCaP cells and VCaP cells (ATCC, LGC standard) were routinely maintained in RPMI 1640 culture medium supplemented with 10% fetal bovine serum (FBS), 5 mg/ml penicillin/streptomycin, and 2 mM l -glutamine (Invitrogen) at 37°C in a humidified atmosphere with 5% CO2 .

    Techniques: In Vitro, In Vivo, Transfection, Western Blot, Cell Culture, Cell Counting

    B20 and PDEP binding of hot ( 3 H-PGE 2 ) and cold (PGE 2 ) in a cell free system. Particles were suspended in RPMI media with 2.5% FBS for 24h, particles were centrifuged and supernatant was collected for standard scintillation counting. B20 at both 100µg/mL

    Journal: Chemosphere

    Article Title: Diesel and biodiesel exhaust particle effects on rat alveolar macrophages with in vitro exposure

    doi: 10.1016/j.chemosphere.2013.10.080

    Figure Lengend Snippet: B20 and PDEP binding of hot ( 3 H-PGE 2 ) and cold (PGE 2 ) in a cell free system. Particles were suspended in RPMI media with 2.5% FBS for 24h, particles were centrifuged and supernatant was collected for standard scintillation counting. B20 at both 100µg/mL

    Article Snippet: The total AMs were diluted at 5 × 105 cells in 1mL of RPMI media (Gibco, Grand Island, NY) with 2.5% FBS (Gibco, Grand Island, NY) and 50µg/mL gentamycin (Gibco, Grand Island, NY).

    Techniques: Binding Assay

    Temsirolimus inhibited VEGF production by cancer cells and endothelial proliferation. ( A ) Tumor cells were incubated with or without HGF (50 ng/ml) in the presence of different concentrations of temsirolimus for 48 h. Then, supernatants were harvested and the number of tumor cells was counted. VEGF concentration in the supernatants was determined by ELISA. VEGF levels corrected by the tumor cell number are shown. ( B ) HMVECs were incubated in RPMI-1640 medium with 10% FBS (control), RPMI-1640 medium with 10%-FBS plus VEGF, or HuMedia-MvG with different concentrations of temsirolimus for 72 h. Then, cell viability was determined using the MTT assay. Bars show SD. The data shown are 2 independent experiments with similar results.

    Journal: PLoS ONE

    Article Title: mTOR Inhibitors Control the Growth of EGFR Mutant Lung Cancer Even after Acquiring Resistance by HGF

    doi: 10.1371/journal.pone.0062104

    Figure Lengend Snippet: Temsirolimus inhibited VEGF production by cancer cells and endothelial proliferation. ( A ) Tumor cells were incubated with or without HGF (50 ng/ml) in the presence of different concentrations of temsirolimus for 48 h. Then, supernatants were harvested and the number of tumor cells was counted. VEGF concentration in the supernatants was determined by ELISA. VEGF levels corrected by the tumor cell number are shown. ( B ) HMVECs were incubated in RPMI-1640 medium with 10% FBS (control), RPMI-1640 medium with 10%-FBS plus VEGF, or HuMedia-MvG with different concentrations of temsirolimus for 72 h. Then, cell viability was determined using the MTT assay. Bars show SD. The data shown are 2 independent experiments with similar results.

    Article Snippet: Tumor cells, plated at 2×103 /100 µL RPMI 1640 medium plus 10% FBS/well in 96-well plates, were incubated for 24 h. Then, erlotinib, temsirolimus, everolimus, and/or HGF were added to each well, and incubation was continued for an additional 72 h. Cell viability was measured with MTT solution (2 mg/mL; Sigma, St. Louis, MO), as previously described .

    Techniques: Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay, MTT Assay

    Activities of antioxidant enzymes in MDA-MB-23 and MCF-7 cells treated with 100 μg/ml of CE. Breast cancer cell lines, MCF-7 and MDA-MB-231, were seeded into 12-well plates containing RPMI 1640 and DMEM, respectively, and supplemented with 10% FBS at 5 × 10 6  cells/well and allowed to attach for 24 h. The cells were treated with 100 μg/ml of CE (IC 70  concentration determined from MTT assay) at varying time points (6, 9, 12, 24, and 48 h incubation). Activity of (A) catalase, (B) superoxide dismutase and (C) glutathione peroxidase was determined using commercial assay kits. Results are expressed as mean ± standard deviation. P

    Journal: PLoS ONE

    Article Title: Cinnamomum cassia Suppresses Caspase-9 through Stimulation of AKT1 in MCF-7 Cells but Not in MDA-MB-231 Cells

    doi: 10.1371/journal.pone.0145216

    Figure Lengend Snippet: Activities of antioxidant enzymes in MDA-MB-23 and MCF-7 cells treated with 100 μg/ml of CE. Breast cancer cell lines, MCF-7 and MDA-MB-231, were seeded into 12-well plates containing RPMI 1640 and DMEM, respectively, and supplemented with 10% FBS at 5 × 10 6 cells/well and allowed to attach for 24 h. The cells were treated with 100 μg/ml of CE (IC 70 concentration determined from MTT assay) at varying time points (6, 9, 12, 24, and 48 h incubation). Activity of (A) catalase, (B) superoxide dismutase and (C) glutathione peroxidase was determined using commercial assay kits. Results are expressed as mean ± standard deviation. P

    Article Snippet: The cell lines were cultured in RPMI-1640 and DMEM (Sigma Aldrich Chemical Company, UK), respectively, and supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin solution at 37°C in a 5% CO2 incubator.

    Techniques: Multiple Displacement Amplification, Concentration Assay, MTT Assay, Incubation, Activity Assay, Standard Deviation

    Actin depolymerization disrupts TEM-HIV-1 polarized assembly platforms. Jurkat LAI cells (5 × 10 5 ) were washed, resuspended in RPMI 1640-1% FCS, and adhered to poly- l -lysine-coated coverslips at 37°C. The cells were either untreated

    Journal: Journal of Virology

    Article Title: Human Immunodeficiency Virus Type 1 Assembly, Budding, and Cell-Cell Spread in T Cells Take Place in Tetraspanin-Enriched Plasma Membrane Domains ▿

    doi: 10.1128/JVI.01845-06

    Figure Lengend Snippet: Actin depolymerization disrupts TEM-HIV-1 polarized assembly platforms. Jurkat LAI cells (5 × 10 5 ) were washed, resuspended in RPMI 1640-1% FCS, and adhered to poly- l -lysine-coated coverslips at 37°C. The cells were either untreated

    Article Snippet: CD4+ T cells (2 × 105 to 5 × 105 ) were washed in RPMI 1640-1% FCS and incubated on poly- l -lysine (Sigma)-treated coverslips at 37°C for up to 60 min.

    Techniques: Transmission Electron Microscopy

    Inhibition of growth by the overexpression of NPRL2. (A) Cell proliferation curves, determined using Cell Counting Kit-8 assays. (B) Colony formation assay, in which 1×10 3 cells were seeded into six-well plates and cultured in RPMI 1640 for 2 weeks. The colonies were then stained with Giemsa, counted and images were captured. Control, cells without transfection; mock, cells transfected with mock vector; NPRL2, cells transfected with NPRL2-overexpression vector. NPRL2, nitrogen permease regulator-like-2.

    Journal: Molecular Medicine Reports

    Article Title: Functional characterization of the nitrogen permease regulator-like-2 candidate tumor suppressor gene in colorectal cancer cell lines

    doi: 10.3892/mmr.2015.3881

    Figure Lengend Snippet: Inhibition of growth by the overexpression of NPRL2. (A) Cell proliferation curves, determined using Cell Counting Kit-8 assays. (B) Colony formation assay, in which 1×10 3 cells were seeded into six-well plates and cultured in RPMI 1640 for 2 weeks. The colonies were then stained with Giemsa, counted and images were captured. Control, cells without transfection; mock, cells transfected with mock vector; NPRL2, cells transfected with NPRL2-overexpression vector. NPRL2, nitrogen permease regulator-like-2.

    Article Snippet: Following culture in RPMI 1640 media, supplemented with 10% FBS at 37°C and 5% CO2 for 2 weeks, the cells were washed twice with phosphate-buffered saline (PBS) and stained with Giemsa (Sigma-Aldrich), following which the number of colonies containing > 50 cells were counted.

    Techniques: Inhibition, Over Expression, Cell Counting, Colony Assay, Cell Culture, Staining, Transfection, Plasmid Preparation

    Altered HIF1α-dependent effects of normoxia and hypoxia under different culture conditions. Western blot analysis was performed 24 h after the application of different treatments for HIF1α and carbonic anhydrase (CAIX), while β-actin served as a control. (Lane 1) Medium control; (lane 2) siRNA against HIF1α; (lane 3) 1% fetal bovine serum; (lane 4) 1% fetal bovine serum and siRNA against HIF1α; (lane 5) 5 mM glutamine; (lane 6) 5 mM glutamine and siRNA against HIF1α; (lane 7) 1% fetal bovine serum and 5 mM glutamine; and, (lane 8) 1% fetal bovine serum, 5 mM glutamine and siRNA against HIF1α under normoxic or hypoxic (*) conditions in MDA-MB-231 cells. The experiments were performed in RPMI 1640 in the presence of 11.1 mM glucose, but without glutamine. Deep sequencing analysis was performed for each of the corresponding RNA samples (lanes 7, 8, 7*, and 8*) from four independent experiments (Please see Supplemental Figure S3 and the results from the densitometric evaluation of the Western blots of four independent experiments).

    Journal: International Journal of Molecular Sciences

    Article Title: Causes and Consequences of A Glutamine Induced Normoxic HIF1 Activity for the Tumor Metabolism

    doi: 10.3390/ijms20194742

    Figure Lengend Snippet: Altered HIF1α-dependent effects of normoxia and hypoxia under different culture conditions. Western blot analysis was performed 24 h after the application of different treatments for HIF1α and carbonic anhydrase (CAIX), while β-actin served as a control. (Lane 1) Medium control; (lane 2) siRNA against HIF1α; (lane 3) 1% fetal bovine serum; (lane 4) 1% fetal bovine serum and siRNA against HIF1α; (lane 5) 5 mM glutamine; (lane 6) 5 mM glutamine and siRNA against HIF1α; (lane 7) 1% fetal bovine serum and 5 mM glutamine; and, (lane 8) 1% fetal bovine serum, 5 mM glutamine and siRNA against HIF1α under normoxic or hypoxic (*) conditions in MDA-MB-231 cells. The experiments were performed in RPMI 1640 in the presence of 11.1 mM glucose, but without glutamine. Deep sequencing analysis was performed for each of the corresponding RNA samples (lanes 7, 8, 7*, and 8*) from four independent experiments (Please see Supplemental Figure S3 and the results from the densitometric evaluation of the Western blots of four independent experiments).

    Article Snippet: Moreover, for experiments, the cells were treated with 3 mM ibuprofen Caesar and Loretz (Caesar and Loretz, Hilden, Germany) dissolved in DMSO or with 10 mM acetylsalicyl acid (dissolved in DMSO (Sigma, Steinheim, Germany) or as Aspirin I.V. 500 mg; Bayer, Leverkusen Germany) under normoxic or hypoxic conditions in RPMI medium containing glutamine, 10% FBS, and 1 mM sodium pyruvate (Invitrogen, Karlsruhe, Germany).

    Techniques: Western Blot, Multiple Displacement Amplification, Sequencing

    Altered HIF1α expression after the application of acetylsalicylic acid or ibuprofen in different cell lines under normoxia. Saos-2, XF354 and MDA-MB-231 cells were treated with 10 mM acetylsalicylic acid or 3 mM ibuprofen in RPMI 1640 medium with 5 mM glutamine under normoxic conditions for 24 h. Western blot analysis was performed with anti-HIF1α monoclonal antibody and anti-β-actin monoclonal antibody as a control. Acetylsalicylic acid or ibuprofen significantly reduced the glutamine-induced expression level of the normoxic HIF1α. Data from three independent experiments (Please see Supplemental Figure S5 and for the densitometric evaluation of the Western blots). For the results of experiments under hypoxic conditions, please see Supplemental Figure S6 .

    Journal: International Journal of Molecular Sciences

    Article Title: Causes and Consequences of A Glutamine Induced Normoxic HIF1 Activity for the Tumor Metabolism

    doi: 10.3390/ijms20194742

    Figure Lengend Snippet: Altered HIF1α expression after the application of acetylsalicylic acid or ibuprofen in different cell lines under normoxia. Saos-2, XF354 and MDA-MB-231 cells were treated with 10 mM acetylsalicylic acid or 3 mM ibuprofen in RPMI 1640 medium with 5 mM glutamine under normoxic conditions for 24 h. Western blot analysis was performed with anti-HIF1α monoclonal antibody and anti-β-actin monoclonal antibody as a control. Acetylsalicylic acid or ibuprofen significantly reduced the glutamine-induced expression level of the normoxic HIF1α. Data from three independent experiments (Please see Supplemental Figure S5 and for the densitometric evaluation of the Western blots). For the results of experiments under hypoxic conditions, please see Supplemental Figure S6 .

    Article Snippet: Moreover, for experiments, the cells were treated with 3 mM ibuprofen Caesar and Loretz (Caesar and Loretz, Hilden, Germany) dissolved in DMSO or with 10 mM acetylsalicyl acid (dissolved in DMSO (Sigma, Steinheim, Germany) or as Aspirin I.V. 500 mg; Bayer, Leverkusen Germany) under normoxic or hypoxic conditions in RPMI medium containing glutamine, 10% FBS, and 1 mM sodium pyruvate (Invitrogen, Karlsruhe, Germany).

    Techniques: Expressing, Multiple Displacement Amplification, Western Blot

    Effects of glutamine application on HIF1α expression in different tumor cell lines. Saos-2, XF354 and MDA-MB-231 cells were left untreated or treated with 5 mM glutamine in RPMI 1640 medium without glutamine under normoxic conditions for 24 h followed by Western blot analysis of HIF1α and β-actin as a loading control. Data from three independent experiments (Please see Supplemental Figure S4 and the results from the densitometric evaluation of the Western blots).

    Journal: International Journal of Molecular Sciences

    Article Title: Causes and Consequences of A Glutamine Induced Normoxic HIF1 Activity for the Tumor Metabolism

    doi: 10.3390/ijms20194742

    Figure Lengend Snippet: Effects of glutamine application on HIF1α expression in different tumor cell lines. Saos-2, XF354 and MDA-MB-231 cells were left untreated or treated with 5 mM glutamine in RPMI 1640 medium without glutamine under normoxic conditions for 24 h followed by Western blot analysis of HIF1α and β-actin as a loading control. Data from three independent experiments (Please see Supplemental Figure S4 and the results from the densitometric evaluation of the Western blots).

    Article Snippet: Moreover, for experiments, the cells were treated with 3 mM ibuprofen Caesar and Loretz (Caesar and Loretz, Hilden, Germany) dissolved in DMSO or with 10 mM acetylsalicyl acid (dissolved in DMSO (Sigma, Steinheim, Germany) or as Aspirin I.V. 500 mg; Bayer, Leverkusen Germany) under normoxic or hypoxic conditions in RPMI medium containing glutamine, 10% FBS, and 1 mM sodium pyruvate (Invitrogen, Karlsruhe, Germany).

    Techniques: Expressing, Multiple Displacement Amplification, Western Blot

    Gene mRNA levels using deep sequencing data of genes involved in glycolysis after HIF1α was knocked down in the MDA-MB-231 cells. RNA was harvested 24 h after the start of the experiments. The experiments were performed in RPMI 1640 medium without glutamine but with 11.1 mM glucose. In this scheme, the mRNA levels are depicted as a ratio of untreated cells to HIF-siRNA transfected cells either under normoxic (N) or under hypoxic (H) conditions. Deep sequencing analysis was performed while using the hypoxic samples after the application of 5 mM L-glutamine and 1% fetal bovine serum (corresponding to lane 7* in Figure 1 ) and hypoxic samples that were additionally treated with HIF1α-specific siRNA (corresponding to lane 8* in Figure 1 ) in four independent experiments. H (hypoxia)-describes the quotients of the transcript level of genes from the hypoxic HIF1-positive samples (corresponding to lane 7* in Figure 1 ) divided by the transcript level from the hypoxic HIF1-negative samples (corresponding to lane 8* in Figure 1 ). N (normoxia) describes quotients of the transcript level of the normoxic HIF1-positive samples (corresponding to lane 7 in Figure 1 ) divided by the transcript level of the normoxic HIF1-negative samples (corresponding to lane 8 in Figure 1 ). Genes marked yellow were significantly upregulated by HIF1 under normoxic conditions, as was validated using qPCR ( Supplemental Figure S16, Supplemental Tables S1 and S2 ). Figure was adapted from [ 34 ].

    Journal: International Journal of Molecular Sciences

    Article Title: Causes and Consequences of A Glutamine Induced Normoxic HIF1 Activity for the Tumor Metabolism

    doi: 10.3390/ijms20194742

    Figure Lengend Snippet: Gene mRNA levels using deep sequencing data of genes involved in glycolysis after HIF1α was knocked down in the MDA-MB-231 cells. RNA was harvested 24 h after the start of the experiments. The experiments were performed in RPMI 1640 medium without glutamine but with 11.1 mM glucose. In this scheme, the mRNA levels are depicted as a ratio of untreated cells to HIF-siRNA transfected cells either under normoxic (N) or under hypoxic (H) conditions. Deep sequencing analysis was performed while using the hypoxic samples after the application of 5 mM L-glutamine and 1% fetal bovine serum (corresponding to lane 7* in Figure 1 ) and hypoxic samples that were additionally treated with HIF1α-specific siRNA (corresponding to lane 8* in Figure 1 ) in four independent experiments. H (hypoxia)-describes the quotients of the transcript level of genes from the hypoxic HIF1-positive samples (corresponding to lane 7* in Figure 1 ) divided by the transcript level from the hypoxic HIF1-negative samples (corresponding to lane 8* in Figure 1 ). N (normoxia) describes quotients of the transcript level of the normoxic HIF1-positive samples (corresponding to lane 7 in Figure 1 ) divided by the transcript level of the normoxic HIF1-negative samples (corresponding to lane 8 in Figure 1 ). Genes marked yellow were significantly upregulated by HIF1 under normoxic conditions, as was validated using qPCR ( Supplemental Figure S16, Supplemental Tables S1 and S2 ). Figure was adapted from [ 34 ].

    Article Snippet: Moreover, for experiments, the cells were treated with 3 mM ibuprofen Caesar and Loretz (Caesar and Loretz, Hilden, Germany) dissolved in DMSO or with 10 mM acetylsalicyl acid (dissolved in DMSO (Sigma, Steinheim, Germany) or as Aspirin I.V. 500 mg; Bayer, Leverkusen Germany) under normoxic or hypoxic conditions in RPMI medium containing glutamine, 10% FBS, and 1 mM sodium pyruvate (Invitrogen, Karlsruhe, Germany).

    Techniques: Sequencing, Multiple Displacement Amplification, Transfection, Real-time Polymerase Chain Reaction

    Glutamine independent induction of normoxic HIF1α via down regulation of ACLY or ACSS2. Application of 20 nM siRNA against ATP citrate lyase (ACLY) or the cytosolic form of the acetyl-CoA synthetase (ACSS2) in Saos-2, XF354 and MDA-MB-231 cells in RPMI 1640 medium without glutamine under normoxic conditions for 48 h. This was followed by Western blot analysis of HIF1α, ACLY (Ser 455), and ACSS2 and β-actin as a loading control. Data from three independent experiments (Please see Supplemental Figure S9a–c and the results from the densitometric evaluation of the Western blots).

    Journal: International Journal of Molecular Sciences

    Article Title: Causes and Consequences of A Glutamine Induced Normoxic HIF1 Activity for the Tumor Metabolism

    doi: 10.3390/ijms20194742

    Figure Lengend Snippet: Glutamine independent induction of normoxic HIF1α via down regulation of ACLY or ACSS2. Application of 20 nM siRNA against ATP citrate lyase (ACLY) or the cytosolic form of the acetyl-CoA synthetase (ACSS2) in Saos-2, XF354 and MDA-MB-231 cells in RPMI 1640 medium without glutamine under normoxic conditions for 48 h. This was followed by Western blot analysis of HIF1α, ACLY (Ser 455), and ACSS2 and β-actin as a loading control. Data from three independent experiments (Please see Supplemental Figure S9a–c and the results from the densitometric evaluation of the Western blots).

    Article Snippet: Moreover, for experiments, the cells were treated with 3 mM ibuprofen Caesar and Loretz (Caesar and Loretz, Hilden, Germany) dissolved in DMSO or with 10 mM acetylsalicyl acid (dissolved in DMSO (Sigma, Steinheim, Germany) or as Aspirin I.V. 500 mg; Bayer, Leverkusen Germany) under normoxic or hypoxic conditions in RPMI medium containing glutamine, 10% FBS, and 1 mM sodium pyruvate (Invitrogen, Karlsruhe, Germany).

    Techniques: Multiple Displacement Amplification, Western Blot

    Altered HIF1α expression after the application of sodium citrate in cells of different lineages under normoxia after simultaneously treatment with glutamine. Saos-2, XF354 and MDA-MB-231 cells were treated with 8 mM (the XF354 cells were treated with only 4 mM sodium citrate) in RPMI 1640 medium with 5 mM glutamine under normoxic conditions for 24 h. Western blot analysis was performed with anti-HIF1α monoclonal antibody and anti-β-actin monoclonal antibody as a control. Sodium citrate significantly reduces the glutamine-induced normoxic HIF1 α-level. Data are from three independent experiments (Please see Supplemental Figure S10 and the results from the densitometric evaluation of the Western blots).

    Journal: International Journal of Molecular Sciences

    Article Title: Causes and Consequences of A Glutamine Induced Normoxic HIF1 Activity for the Tumor Metabolism

    doi: 10.3390/ijms20194742

    Figure Lengend Snippet: Altered HIF1α expression after the application of sodium citrate in cells of different lineages under normoxia after simultaneously treatment with glutamine. Saos-2, XF354 and MDA-MB-231 cells were treated with 8 mM (the XF354 cells were treated with only 4 mM sodium citrate) in RPMI 1640 medium with 5 mM glutamine under normoxic conditions for 24 h. Western blot analysis was performed with anti-HIF1α monoclonal antibody and anti-β-actin monoclonal antibody as a control. Sodium citrate significantly reduces the glutamine-induced normoxic HIF1 α-level. Data are from three independent experiments (Please see Supplemental Figure S10 and the results from the densitometric evaluation of the Western blots).

    Article Snippet: Moreover, for experiments, the cells were treated with 3 mM ibuprofen Caesar and Loretz (Caesar and Loretz, Hilden, Germany) dissolved in DMSO or with 10 mM acetylsalicyl acid (dissolved in DMSO (Sigma, Steinheim, Germany) or as Aspirin I.V. 500 mg; Bayer, Leverkusen Germany) under normoxic or hypoxic conditions in RPMI medium containing glutamine, 10% FBS, and 1 mM sodium pyruvate (Invitrogen, Karlsruhe, Germany).

    Techniques: Expressing, Multiple Displacement Amplification, Western Blot

    (A) LDH release from MCF-7 cells subjected to 2 μmol/L fluopsin C treatment. MCF-7 cells were treated with 1% Triton-X-100 (Sigma) in RPMI-1640 medium for 10 min as the 100% LDH release control. b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Fluopsin C induces oncosis of human breast adenocarcinoma cells

    doi: 10.1038/aps.2013.44

    Figure Lengend Snippet: (A) LDH release from MCF-7 cells subjected to 2 μmol/L fluopsin C treatment. MCF-7 cells were treated with 1% Triton-X-100 (Sigma) in RPMI-1640 medium for 10 min as the 100% LDH release control. b P

    Article Snippet: The MCF-7 and HL7702 cells were maintained in RPMI-1640 medium (Hyclone, Thermo Fisher Scientific, Beijing, China), and the MD-MBA-231 cells were cultured in modified DMEM (Gibco, Invitrogen Corporation, Grand Island, NY, USA).

    Techniques:

    Interaction of ghA, ghB, and ghC with monocyte/macrophages . PMBCs (1 × 10 6 ) were seeded on 13 mm coverslips and incubated in complete RPMI 1640 medium for 2 weeks at 37°C in 5% CO 2 incubator. Cells were treated with 10 μg of each globular head module and incubated with serum-free RPMI 1640 medium for 1 h at 37°C. After washing with PBS, cells were fixed with 4% PFA, permeabilzed with Triton X-100, and probed with anti-gC1qR polyclonal antibody and anti-MBP monoclonal antibody to reveal gC1qR and bound globular head modules, respectively. Cells were washed and treated with Alexa Fluor 488 conjugated secondary goat anti-rabbit antibody and Alexa Fluor 647 conjugated secondary donkey anti-mouse antibody and the nucleus was stained with Hoechst 33342. Cells were then examined under Leica fluorescence microscope with 40× magnification. In the merged images gC1qR is green; globular heads are red; and nucleus is blue. Arrows point to bound globular heads with colocalization of globular heads and gC1qR seen in orange in the merged images. Scale bars: 10 μm.

    Journal: Frontiers in Immunology

    Article Title: Analysis of the Interaction between Globular Head Modules of Human C1q and Its Candidate Receptor gC1qR

    doi: 10.3389/fimmu.2016.00567

    Figure Lengend Snippet: Interaction of ghA, ghB, and ghC with monocyte/macrophages . PMBCs (1 × 10 6 ) were seeded on 13 mm coverslips and incubated in complete RPMI 1640 medium for 2 weeks at 37°C in 5% CO 2 incubator. Cells were treated with 10 μg of each globular head module and incubated with serum-free RPMI 1640 medium for 1 h at 37°C. After washing with PBS, cells were fixed with 4% PFA, permeabilzed with Triton X-100, and probed with anti-gC1qR polyclonal antibody and anti-MBP monoclonal antibody to reveal gC1qR and bound globular head modules, respectively. Cells were washed and treated with Alexa Fluor 488 conjugated secondary goat anti-rabbit antibody and Alexa Fluor 647 conjugated secondary donkey anti-mouse antibody and the nucleus was stained with Hoechst 33342. Cells were then examined under Leica fluorescence microscope with 40× magnification. In the merged images gC1qR is green; globular heads are red; and nucleus is blue. Arrows point to bound globular heads with colocalization of globular heads and gC1qR seen in orange in the merged images. Scale bars: 10 μm.

    Article Snippet: To isolate monocytes, blood in RPMI 1640 was separated on a Ficol column (Ficol-Plaque Plus, GE healthcare) by centrifugation at 2000 rpm for 16 min at room temperature.

    Techniques: Incubation, Staining, Fluorescence, Microscopy

    Anti-tumor effect of G47Δ in NPC xenografts in mice. CNE-2 cells (1 × 10 6 ) and SUNE-1 cells (5 × 10 5 ) were suspended in 100 mL RPMI-1640 complete medium with 25% Matrigel (BD Biosciences) and implanted subcutaneous into the left flanks of 4-week-old nude mice. When the maximal diameter of the subcutaneous tumors was approximately 5 mm, G47Δ (2 × 10 7 pfu /50 µL) or virus buffer (150 mmol/L NaCl and 20 mmol/L Tris at pH7.5) was injected into the subcutaneous tumors twice a week for 2 weeks. A, the subcutaneous CNE-2 tumor regressed by treatment with G47Δ on day 34 or appeared as large tumors after the mock treatment on day 20 post-injection of tumor cells. B, the subcutaneous SUNE-1 tumor regressed by treatment with G47Δ or appeared as large tumors after the mock treatment on day 37 post-injection of tumor cells. Comparison of mice with subcutaneous CNE-2 tumors (C) or subcutaneous SUNE-1 tumors (D) in the mock-treated group (upper) and the virus-treated group (lower) on day 20 or day 37 post-injection of tumor cells. Hematoxylin and eosin (HE) staining of CNE-2 (E), SUNE-1 (F), G47Δ-treated CNE-2 (G), and G47Δ-treated SUNE-1 (H) subcutaneous tumors (×200).

    Journal: Chinese Journal of Cancer

    Article Title: Anti-tumor effect of oncolytic herpes simplex virus G47delta on human nasopharyngeal carcinoma

    doi: 10.5732/cjc.011.10301

    Figure Lengend Snippet: Anti-tumor effect of G47Δ in NPC xenografts in mice. CNE-2 cells (1 × 10 6 ) and SUNE-1 cells (5 × 10 5 ) were suspended in 100 mL RPMI-1640 complete medium with 25% Matrigel (BD Biosciences) and implanted subcutaneous into the left flanks of 4-week-old nude mice. When the maximal diameter of the subcutaneous tumors was approximately 5 mm, G47Δ (2 × 10 7 pfu /50 µL) or virus buffer (150 mmol/L NaCl and 20 mmol/L Tris at pH7.5) was injected into the subcutaneous tumors twice a week for 2 weeks. A, the subcutaneous CNE-2 tumor regressed by treatment with G47Δ on day 34 or appeared as large tumors after the mock treatment on day 20 post-injection of tumor cells. B, the subcutaneous SUNE-1 tumor regressed by treatment with G47Δ or appeared as large tumors after the mock treatment on day 37 post-injection of tumor cells. Comparison of mice with subcutaneous CNE-2 tumors (C) or subcutaneous SUNE-1 tumors (D) in the mock-treated group (upper) and the virus-treated group (lower) on day 20 or day 37 post-injection of tumor cells. Hematoxylin and eosin (HE) staining of CNE-2 (E), SUNE-1 (F), G47Δ-treated CNE-2 (G), and G47Δ-treated SUNE-1 (H) subcutaneous tumors (×200).

    Article Snippet: Cells and viruses Two poorly differentiated NPC cell lines, CNE-2 and SUNE-1, were cultured in RPMI-1640 with 4.5 g/L glucose (Mediatech, Inc., Herndon, VA) supplemented with 10% fetal calf serum (Hyclone Laboratories, Logan, UT) at 37°C in an atmosphere of 5% CO2 .

    Techniques: Mouse Assay, Injection, Staining

    Scanning electron photomicrographs of METs formed by bovine macrophages in response to M. haemolytica cells. Bovine macrophages (2.5 × 10 5 ) were incubated with 5 × 10 7 M. haemolytica cells or RPMI 1640 (negative control) for 5 min at 37°C.

    Journal: Infection and Immunity

    Article Title: Mannheimia haemolytica and Its Leukotoxin Cause Macrophage Extracellular Trap Formation by Bovine Macrophages

    doi: 10.1128/IAI.06120-11

    Figure Lengend Snippet: Scanning electron photomicrographs of METs formed by bovine macrophages in response to M. haemolytica cells. Bovine macrophages (2.5 × 10 5 ) were incubated with 5 × 10 7 M. haemolytica cells or RPMI 1640 (negative control) for 5 min at 37°C.

    Article Snippet: Adherent monocytes were allowed to differentiate into monocyte-derived macrophages by incubating them in RPMI 1640 with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin for 7 days at 37°C.

    Techniques: Incubation, Negative Control

    IMPDH forms filaments during ex vivo primary human T cell activation. (A) Representative images of T cells left untreated or stimulated by mitogens PHA, ConA, or anti-CD3/CD28 for 72 h, then fixed and stained for IMPDH (green) and T cell marker CD3 (red). White arrows: examples of IMPDH filaments. Yellow arrows: example of a ring-shaped filament. Images were captured using identical microscope settings for all treatment groups. (B) Quantification of the percentage of T cells that form filaments when untreated or treated with mitogens PHA, ConA, or anti-CD3/CD28. Cells were cultured in RPMI 1640 under four different conditions: 2 mM glutamine (Gln), 2 mM Gln + 1 h fresh medium, 16 mM Gln, or 16 mM Gln + 1 h fresh medium (represented by differently shaded bars). Different culture conditions were grouped together according to mitogenic treatment and compared to untreated cells (e.g., all PHA-treated conditions grouped vs. all untreated conditions grouped) and statistical significance displayed above each group. No significant differences were observed within these treatment groups due to culture conditions (e.g., PHA 2 mM Gln vs. PHA 16 mM Gln, no difference). Statistical test used: two-way ANOVA followed by Tukey's multiple comparisons test; **** p

    Journal: Frontiers in Immunology

    Article Title: Immune Response-Dependent Assembly of IMP Dehydrogenase Filaments

    doi: 10.3389/fimmu.2018.02789

    Figure Lengend Snippet: IMPDH forms filaments during ex vivo primary human T cell activation. (A) Representative images of T cells left untreated or stimulated by mitogens PHA, ConA, or anti-CD3/CD28 for 72 h, then fixed and stained for IMPDH (green) and T cell marker CD3 (red). White arrows: examples of IMPDH filaments. Yellow arrows: example of a ring-shaped filament. Images were captured using identical microscope settings for all treatment groups. (B) Quantification of the percentage of T cells that form filaments when untreated or treated with mitogens PHA, ConA, or anti-CD3/CD28. Cells were cultured in RPMI 1640 under four different conditions: 2 mM glutamine (Gln), 2 mM Gln + 1 h fresh medium, 16 mM Gln, or 16 mM Gln + 1 h fresh medium (represented by differently shaded bars). Different culture conditions were grouped together according to mitogenic treatment and compared to untreated cells (e.g., all PHA-treated conditions grouped vs. all untreated conditions grouped) and statistical significance displayed above each group. No significant differences were observed within these treatment groups due to culture conditions (e.g., PHA 2 mM Gln vs. PHA 16 mM Gln, no difference). Statistical test used: two-way ANOVA followed by Tukey's multiple comparisons test; **** p

    Article Snippet: PBMCs were stimulated with PHA, ConA, or anti-CD3/CD28 and harvested after culture in RPMI 1640 for 72 h under 4 different conditions: (1) 2 mM glutamine, (2) 2 mM glutamine + 1 h fresh medium, (3) 16 mM glutamine, and (4) 16 mM glutamine + 1 h fresh medium.

    Techniques: Ex Vivo, Activation Assay, Staining, Marker, Microscopy, Cell Culture

    Compound 6-B345TTQ increases α4-dependent cell spreading in Jurkat T cells and CHO cells. Jurkat T cells expressing wild-type α4 ( A ) or α4(S988A) integrin-expressing CHO cells ( B ) were resuspended in RPMI 1640 medium or Dulbecco's

    Journal: The Journal of Biological Chemistry

    Article Title: A Small Molecule That Inhibits the Interaction of Paxillin and ?4 Integrin Inhibits Accumulation of Mononuclear Leukocytes at a Site of Inflammation *

    doi: 10.1074/jbc.M109.066993

    Figure Lengend Snippet: Compound 6-B345TTQ increases α4-dependent cell spreading in Jurkat T cells and CHO cells. Jurkat T cells expressing wild-type α4 ( A ) or α4(S988A) integrin-expressing CHO cells ( B ) were resuspended in RPMI 1640 medium or Dulbecco's

    Article Snippet: THP-1 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 10 m m Hepes, 50 μg/ml gentamycin, 50 μ m 2-mercaptoethanol, and 50 μg/ml gentamycin.

    Techniques: Expressing

    Foam cell formation in hMDMs. Similar to oxLDL uptake studies, ether-linked AMs ( 3 ) displayed significantly higher efficacy than ester-linked AMs ( 5 ) at preventing the foam cell phenotype from developing, minimizing intracellular lipid accumulation. hMDMs were incubated with 50 µg/mL oxLDL and 10 −5 M AMs in RPMI 1640 for 24 h before staining with Oil Red O and Hoechst 33342.

    Journal: Biomaterials

    Article Title: Tartaric Acid-based Amphiphilic Macromolecules with Ether Linkages Exhibit Enhanced Repression of Oxidized Low Density Lipoprotein Uptake

    doi: 10.1016/j.biomaterials.2015.02.038

    Figure Lengend Snippet: Foam cell formation in hMDMs. Similar to oxLDL uptake studies, ether-linked AMs ( 3 ) displayed significantly higher efficacy than ester-linked AMs ( 5 ) at preventing the foam cell phenotype from developing, minimizing intracellular lipid accumulation. hMDMs were incubated with 50 µg/mL oxLDL and 10 −5 M AMs in RPMI 1640 for 24 h before staining with Oil Red O and Hoechst 33342.

    Article Snippet: To measure foam cell formation, hMDMs were incubated with 50 µg/mL oxLDL and 10−5 M AMs in RPMI 1640 for 24 hours before fixation in 4% paraformaldehyde.

    Techniques: Affinity Magnetic Separation, Incubation, Staining

    oxLDL uptake inhibition by AMs in hMDMs. All ether-linked AMs had significant reductions in oxLDL uptake, with increased efficacy being directly correlated to longer aliphatic chains. hMDMs were incubated with 5 µg/mL oxLDL and 10 −6 M AMs in RPMI 1640 for 24 h then analyzed via flow cytometry. * corresponds to p

    Journal: Biomaterials

    Article Title: Tartaric Acid-based Amphiphilic Macromolecules with Ether Linkages Exhibit Enhanced Repression of Oxidized Low Density Lipoprotein Uptake

    doi: 10.1016/j.biomaterials.2015.02.038

    Figure Lengend Snippet: oxLDL uptake inhibition by AMs in hMDMs. All ether-linked AMs had significant reductions in oxLDL uptake, with increased efficacy being directly correlated to longer aliphatic chains. hMDMs were incubated with 5 µg/mL oxLDL and 10 −6 M AMs in RPMI 1640 for 24 h then analyzed via flow cytometry. * corresponds to p

    Article Snippet: To measure foam cell formation, hMDMs were incubated with 50 µg/mL oxLDL and 10−5 M AMs in RPMI 1640 for 24 hours before fixation in 4% paraformaldehyde.

    Techniques: Inhibition, Affinity Magnetic Separation, Incubation, Flow Cytometry, Cytometry