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Corning Life Sciences fetal bovine serum fbs
Fetal Bovine Serum Fbs, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/fetal bovine serum fbs/product/Corning Life Sciences
Average 86 stars, based on 1 article reviews
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fetal bovine serum fbs - by Bioz Stars, 2021-04
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Injection:

Article Title: Regulation of Head and Neck Squamous Cancer Stem Cells by PI3K and SOX2
Article Snippet: .. CSCs were sorted into PBS +2% fetal bovine serum (FBS), suspended in 1:1, DMEM +10% FBS:Matrigel (Corning, Corning, NY) to 100 µL per injection, and injected (25 g needle) into the flanks of nude mice. ..

Mouse Assay:

Article Title: Regulation of Head and Neck Squamous Cancer Stem Cells by PI3K and SOX2
Article Snippet: .. CSCs were sorted into PBS +2% fetal bovine serum (FBS), suspended in 1:1, DMEM +10% FBS:Matrigel (Corning, Corning, NY) to 100 µL per injection, and injected (25 g needle) into the flanks of nude mice. ..

Article Title: Targeting cellular heterogeneity with CXCR2 blockade for the treatment of therapy-resistant prostate cancer
Article Snippet: Immunocompromised NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl /SzJ) and nude (nu/nu) mice were from The Jackson Laboratories. .. A total of 2 × 106 of LNCaP or LNCaP-CXCR2 cells or 1 × 106 of C4-2B/MDVR, C4-2B, or PC3 cells were suspended in 0.1-ml 1× RPMI 1640 and 10% fetal bovine serum (FBS) with 50% Matrigel (Corning), then inoculated subcutaneously into the bilateral flanks of six-week-old male NSG (LNCaP LNCaP-CXCR2, C4-2B, and C4-2B/MDVR) nude (PC3) mice, as previously described ( ). .. For LNCaP and LNCaP-CXCR2 xenografts, mice were euthanized with CO2 12 weeks after implantation, and the xenografts were harvested, fixed in 10% formalin, and embedded in paraffin for subsequent analysis.

Cell Culture:

Article Title: Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3
Article Snippet: Thus, either loss of Fbxl8 or gain of a cyclin D3 mutation that stabilizes cyclin D3 protein level will accelerate cell cycle progression and lymphoma cell growth consistent Fbxl8 functioning as a tumor suppressor (Fig. ). .. Cell culture, transfection, infection, cell cycle analyses and CRISPR-Cas9 knockoutNIH3T3 and 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning). .. U2OS cells were cultured in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning).

Article Title: PD-L1 chimeric costimulatory receptor improves the efficacy of CAR-T cells for PD-L1-positive solid tumors and reduces toxicity in vivo
Article Snippet: Cell lines The following cell lines were used: K562 (a chronic myelogenous leukemia cell line, ATCC #CCL-243), Jurkat, clone E6–1 (an acute T cell leukemia cell line, ATCC #TIB-152), A549 (a lung cancer cell line, ATCC #CCL-185), NCI-H292 (a lung cancer cell line, ATCC #CRL-1848), SKOV3 (an ovarian cancer cell line, HTB-77) and HEK293T (an embryonic kidney cell line, ATCC #CRL-3216). .. All cell lines were purchased from ATCC and maintained in RPMI 1640 (Corning #10–040-CVR) supplemented with 10% fetal bovine serum (FBS) (BI #04–001-1acs) and 1% penicillin-streptomycin (Corning #30–002-CI), except for HEK293T cells, which were cultured in DMEM (Corning #10–013-CV) supplemented with 10% FBS and 1% penicillin-streptomycin. ..

Transfection:

Article Title: Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3
Article Snippet: Thus, either loss of Fbxl8 or gain of a cyclin D3 mutation that stabilizes cyclin D3 protein level will accelerate cell cycle progression and lymphoma cell growth consistent Fbxl8 functioning as a tumor suppressor (Fig. ). .. Cell culture, transfection, infection, cell cycle analyses and CRISPR-Cas9 knockoutNIH3T3 and 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning). .. U2OS cells were cultured in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning).

Infection:

Article Title: Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3
Article Snippet: Thus, either loss of Fbxl8 or gain of a cyclin D3 mutation that stabilizes cyclin D3 protein level will accelerate cell cycle progression and lymphoma cell growth consistent Fbxl8 functioning as a tumor suppressor (Fig. ). .. Cell culture, transfection, infection, cell cycle analyses and CRISPR-Cas9 knockoutNIH3T3 and 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning). .. U2OS cells were cultured in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning).

CRISPR:

Article Title: Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3
Article Snippet: Thus, either loss of Fbxl8 or gain of a cyclin D3 mutation that stabilizes cyclin D3 protein level will accelerate cell cycle progression and lymphoma cell growth consistent Fbxl8 functioning as a tumor suppressor (Fig. ). .. Cell culture, transfection, infection, cell cycle analyses and CRISPR-Cas9 knockoutNIH3T3 and 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning). .. U2OS cells were cultured in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning).

Modification:

Article Title: Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3
Article Snippet: Thus, either loss of Fbxl8 or gain of a cyclin D3 mutation that stabilizes cyclin D3 protein level will accelerate cell cycle progression and lymphoma cell growth consistent Fbxl8 functioning as a tumor suppressor (Fig. ). .. Cell culture, transfection, infection, cell cycle analyses and CRISPR-Cas9 knockoutNIH3T3 and 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning). .. U2OS cells were cultured in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning).

Article Title: Combination of Inhibitors of USP7 and PLK1 has a Strong Synergism against Paclitaxel Resistance
Article Snippet: .. MaterialsDulbecco’s modified Eagle’s medium (DMEM), Minimum essential medium (MEM), Roswell Park Memorial Institute (RPMI)-1640 medium, fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Corning Cellgro (Manassas, VA, USA). .. Volasertib and P22077 were from Selleckchem (Houston, TX, USA) and Lifesensors (Malvern, PA, USA), respectively.

Article Title: Qishen granules inhibit myocardial inflammation injury through regulating arachidonic acid metabolism
Article Snippet: .. Materials Dulbecco Modified Eagle Mediumas (DMEM) supplemented with 10% Fetal Bovine Serum (FBS), Penicillin (100 U/mL) and Streptomycin (100 μg/mL) was purchased from Corning (New York, USA). .. LPS was purchased from Sigma Chemical Co. (St. Louis, USA).

other:

Article Title: Rapid Determination of Plasmonic Nanoparticle Agglomeration Status in Blood
Article Snippet: Phosphate buffered saline (PBS), fetal bovine serum (FBS), and Eagle medium were acquired from Corning Cellgro (Manassas, VA).

Cell Migration Assay:

Article Title: Inhibition of intimal hyperplasia in murine aortic allografts by administration of a small-molecule TLR4 inhibitor TAK-242
Article Snippet: .. Cell migration assay Transwell assay was performed to evaluate VSMC migration in DMEM supplemented with 10% fetal bovine serum (FBS) in the presence or absence of macrophages in a transwell chamber (8-μm pore size; Corning). .. Briefly, macrophages were seeded into the lower chamber in DMEM medium containing 10% FBS and TAK-242, or CCL2 antibody (100 ng/ml, Santa Cruz, Santa Cruz, CA) that neutralized macrophage-induced migration.

Transwell Assay:

Article Title: Inhibition of intimal hyperplasia in murine aortic allografts by administration of a small-molecule TLR4 inhibitor TAK-242
Article Snippet: .. Cell migration assay Transwell assay was performed to evaluate VSMC migration in DMEM supplemented with 10% fetal bovine serum (FBS) in the presence or absence of macrophages in a transwell chamber (8-μm pore size; Corning). .. Briefly, macrophages were seeded into the lower chamber in DMEM medium containing 10% FBS and TAK-242, or CCL2 antibody (100 ng/ml, Santa Cruz, Santa Cruz, CA) that neutralized macrophage-induced migration.

Migration:

Article Title: Inhibition of intimal hyperplasia in murine aortic allografts by administration of a small-molecule TLR4 inhibitor TAK-242
Article Snippet: .. Cell migration assay Transwell assay was performed to evaluate VSMC migration in DMEM supplemented with 10% fetal bovine serum (FBS) in the presence or absence of macrophages in a transwell chamber (8-μm pore size; Corning). .. Briefly, macrophages were seeded into the lower chamber in DMEM medium containing 10% FBS and TAK-242, or CCL2 antibody (100 ng/ml, Santa Cruz, Santa Cruz, CA) that neutralized macrophage-induced migration.

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  • 86
    Corning Life Sciences transwell upper chamber
    RA inhibited dendritic cell (DC) activation and T cell priming in vivo Mice were i.p. injected with RA (250 μg) or DMSO (control) daily from -3 to 1 dpi. These animals were i.v. injected with 3 × 10 9 pfu of AdLacZ and sacrificed at 2 dpi. (A) DCs in the spleen and liver were first gated on CD11b + CD11c + cells and the mean fluorescence intensity (MFI) of co-stimulatory molecules was examined. (B) Bone marrow-derived DCs were generated from B6 mice by using complete RPMI 1640 with 20 ng/ml rGM-CSF. RA (1 μM) was added at day 3 of an 8-day DC culture. Upper panel : Surface CD11c expression level of control and RA-treated DCs. Grey line: control DCs; black line: RA-treated DCs. Lower panel : MFI of CD11c on DCs. (C) RA-treated and control DCs were infected with Ad-LacZ (MOI: 300) for 12 h. Surface molecules were analyzed by flow cytometry. Shaded: Isotype control; grey line: uninfected DCs; black line: Ad-infected DCs. (D) 2 × 10 5 control or RA-treated DCs were cultured, respectively, in the upper chambers of 24-well <t>transwell</t> plates. The lower chambers contained complete medium with rCCL21 (100 ng/ml). After 8 h of culture, the cells in the lower chambers were collected for photographing and counting. (E) Virus-exposed DCs or naïve DCs (5 × 10 5 in 20 μl of PBS) were injected s.c. into the hind footpad of naïve mice. Three types of DC were transferred: naïve DCs, virus-exposed control DCs, and virus-exposed RA-treated DCs ( prepared as in Fig 5B and C). Popliteal draining lymph nodes (LNs) were harvested at 7 dpi. Lymphocytes were isolated and stimulated by PMA and Ionomycin plus Golgistop for 4 h. Percentages of cytokine-producing CD4 + and CD8 + T cells in draining LNs were examined by flow cytometry. Each experiment was repeated at least three times independently. Shown are representative flow cytometric results. * p
    Transwell Upper Chamber, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transwell upper chamber/product/Corning Life Sciences
    Average 86 stars, based on 1 article reviews
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    86
    Corning Life Sciences cd4 t cells
    IL-6 stimulation increases productive infection in resting CD4 +  T cells without T cell activation.  (A)  Three different cytokines, IL-6, IL-8, and CCL2, were added to resting CD4 +  T cells 1 day before infection with a virus expressing GFP. Cytokines were replenished every other day after infection, and GFP-positive rates were measured on day 7 postinfection.  (B)  IL-6 concentration is positively associated with infection rates. Resting T cells were cultured with various concentrations of IL-6 1 day before infection with a reporter virus. The cytokine was replenished on days 1, 3, and 5 postinfection, and GFP was measured on day 6 postinfection. Resting T cells alone and resting T cells cultured with EC −  or EC +  were used as controls.  (C)  EC stimulation is blocked by anti-IL-6 antibody. An anti-IL6 antibody (Ab) was added to EC +/−  for approximately an hour before addition of resting T cells in direct contact at three concentrations (10, 5, and 2 μg/ml). After culturing for a day, the cells were infected with a virus expressing GFP. The antibodies were refreshed on days 1 and 3 postinfection, and GFP levels were measured on day 6 postinfection.  (D)  Similar to the experiment in  (C) , except that T cells were cultured in transwell with EC. Antibody level was at 10 μg/ml. Resting CD4 +  T cells cultured alone or treated with IL-6 served as controls.  (E)  Lack of T cell activation by IL-6 stimulation. Resting CD4 +  T cells were cocultured with IL-6 at 1 ng/ml for 1 day and then infected with a GFP reporter virus. CD25, CD69, and HLA-DR levels were measured on the day of infection (D0) and day 3 (D3) and 6 (D6) postinfection. The cytokine was replenished on days 1, 3, and 5 postinfection, and GFP was measured on day 6 postinfection (the  right  two bars). Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of at least three independent experiments yielding similar results. *Student  t -test;  p
    Cd4 T Cells, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd4 t cells/product/Corning Life Sciences
    Average 86 stars, based on 1 article reviews
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    98
    Corning Life Sciences fetal bovine serum fbs
    Overexpression of Fbxl8 recedes G1-S phase transition of cell cycle through degradation of cyclin D3. a Schematic model of experiment. NIH3T3 cells were transfected with MigR1 IRES-GFP, MigR1Fbxl8 IRES-GFP or MigR1Fbxl8ΔF IRES-GFP, and arrested at G0/G1 phase by serum starvation for 36 h. GFP positive cells and negative cells were FACS sorted and S phase entry was assessed by BrdU incorporation (30 min). b Western analysis of lysates from sorted GFP positive NIH3T3 cells expressing GFP, Fbxl8, or Fbxl8ΔF for cyclin D3, cyclin D1, Flag-Fbxl8 and βactin. The numbers indicate quantifications of cyclin D3 and cyclin D1 normalized by βactin. c GFP and BrdU double-positive cells were analyzed 9, 12, 15, 18, 21 h after release from G0/G1 phase by re-splitting cells in <t>DMEM</t> with <t>10%FBS.</t> Quantification of GFP and BrdU double-positive cells; mean ± SD, * p
    Fetal Bovine Serum Fbs, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fetal bovine serum fbs/product/Corning Life Sciences
    Average 98 stars, based on 1 article reviews
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    99
    Corning Life Sciences transwell apparatus
    Silence of FOXO1 mitigates the inhibitory effects of miR‐5188 knockdown on glioma cell proliferation, invasion and migration. MTT assays (A), EdU incorporation assays (B), flow cytometry assays (C), <t>transwell</t> assays (D) and Boyden assays (E) were performed in U251 or U87 cells treated with control sequence, FOXO1 siRNA or miR‐5188 inhibitor, as indicated. Scale bars, 100 μm. F, Western blotting analyses were applied to measure the protein levels of FOXO1, P‐PI3K, PI3K, P‐AKT, AKT, CCND1, CDK4, c‐JUN, N‐cadherin and vimentin in U251 or U87 cells treated with control sequence, FOXO1 siRNA or miR‐5188 inhibitor, as indicated. G, Western blotting analysis analyses were applied to measure the protein levels of FOXO1, P‐PI3K, PI3K, P‐AKT, AKT, CCND1, CDK4, c‐JUN, N‐cadherin and vimentin protein levels in U87 and U251 cells transfected with control sequence or FOXO1 siRNA. * P
    Transwell Apparatus, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    RA inhibited dendritic cell (DC) activation and T cell priming in vivo Mice were i.p. injected with RA (250 μg) or DMSO (control) daily from -3 to 1 dpi. These animals were i.v. injected with 3 × 10 9 pfu of AdLacZ and sacrificed at 2 dpi. (A) DCs in the spleen and liver were first gated on CD11b + CD11c + cells and the mean fluorescence intensity (MFI) of co-stimulatory molecules was examined. (B) Bone marrow-derived DCs were generated from B6 mice by using complete RPMI 1640 with 20 ng/ml rGM-CSF. RA (1 μM) was added at day 3 of an 8-day DC culture. Upper panel : Surface CD11c expression level of control and RA-treated DCs. Grey line: control DCs; black line: RA-treated DCs. Lower panel : MFI of CD11c on DCs. (C) RA-treated and control DCs were infected with Ad-LacZ (MOI: 300) for 12 h. Surface molecules were analyzed by flow cytometry. Shaded: Isotype control; grey line: uninfected DCs; black line: Ad-infected DCs. (D) 2 × 10 5 control or RA-treated DCs were cultured, respectively, in the upper chambers of 24-well transwell plates. The lower chambers contained complete medium with rCCL21 (100 ng/ml). After 8 h of culture, the cells in the lower chambers were collected for photographing and counting. (E) Virus-exposed DCs or naïve DCs (5 × 10 5 in 20 μl of PBS) were injected s.c. into the hind footpad of naïve mice. Three types of DC were transferred: naïve DCs, virus-exposed control DCs, and virus-exposed RA-treated DCs ( prepared as in Fig 5B and C). Popliteal draining lymph nodes (LNs) were harvested at 7 dpi. Lymphocytes were isolated and stimulated by PMA and Ionomycin plus Golgistop for 4 h. Percentages of cytokine-producing CD4 + and CD8 + T cells in draining LNs were examined by flow cytometry. Each experiment was repeated at least three times independently. Shown are representative flow cytometric results. * p

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

    Article Title: Retinoic acid regulates immune responses by promoting IL-22 and modulating S100 proteins in viral hepatitis

    doi: 10.4049/jimmunol.1601891

    Figure Lengend Snippet: RA inhibited dendritic cell (DC) activation and T cell priming in vivo Mice were i.p. injected with RA (250 μg) or DMSO (control) daily from -3 to 1 dpi. These animals were i.v. injected with 3 × 10 9 pfu of AdLacZ and sacrificed at 2 dpi. (A) DCs in the spleen and liver were first gated on CD11b + CD11c + cells and the mean fluorescence intensity (MFI) of co-stimulatory molecules was examined. (B) Bone marrow-derived DCs were generated from B6 mice by using complete RPMI 1640 with 20 ng/ml rGM-CSF. RA (1 μM) was added at day 3 of an 8-day DC culture. Upper panel : Surface CD11c expression level of control and RA-treated DCs. Grey line: control DCs; black line: RA-treated DCs. Lower panel : MFI of CD11c on DCs. (C) RA-treated and control DCs were infected with Ad-LacZ (MOI: 300) for 12 h. Surface molecules were analyzed by flow cytometry. Shaded: Isotype control; grey line: uninfected DCs; black line: Ad-infected DCs. (D) 2 × 10 5 control or RA-treated DCs were cultured, respectively, in the upper chambers of 24-well transwell plates. The lower chambers contained complete medium with rCCL21 (100 ng/ml). After 8 h of culture, the cells in the lower chambers were collected for photographing and counting. (E) Virus-exposed DCs or naïve DCs (5 × 10 5 in 20 μl of PBS) were injected s.c. into the hind footpad of naïve mice. Three types of DC were transferred: naïve DCs, virus-exposed control DCs, and virus-exposed RA-treated DCs ( prepared as in Fig 5B and C). Popliteal draining lymph nodes (LNs) were harvested at 7 dpi. Lymphocytes were isolated and stimulated by PMA and Ionomycin plus Golgistop for 4 h. Percentages of cytokine-producing CD4 + and CD8 + T cells in draining LNs were examined by flow cytometry. Each experiment was repeated at least three times independently. Shown are representative flow cytometric results. * p

    Article Snippet: DC (2 × 105 ) were re-suspended in complete medium with 2% FBS and placed in a Transwell upper chamber (Costar 3422, Corning).

    Techniques: Activation Assay, In Vivo, Mouse Assay, Injection, Fluorescence, Derivative Assay, Generated, Expressing, Infection, Flow Cytometry, Cytometry, Cell Culture, Isolation

    IL-6 stimulation increases productive infection in resting CD4 +  T cells without T cell activation.  (A)  Three different cytokines, IL-6, IL-8, and CCL2, were added to resting CD4 +  T cells 1 day before infection with a virus expressing GFP. Cytokines were replenished every other day after infection, and GFP-positive rates were measured on day 7 postinfection.  (B)  IL-6 concentration is positively associated with infection rates. Resting T cells were cultured with various concentrations of IL-6 1 day before infection with a reporter virus. The cytokine was replenished on days 1, 3, and 5 postinfection, and GFP was measured on day 6 postinfection. Resting T cells alone and resting T cells cultured with EC −  or EC +  were used as controls.  (C)  EC stimulation is blocked by anti-IL-6 antibody. An anti-IL6 antibody (Ab) was added to EC +/−  for approximately an hour before addition of resting T cells in direct contact at three concentrations (10, 5, and 2 μg/ml). After culturing for a day, the cells were infected with a virus expressing GFP. The antibodies were refreshed on days 1 and 3 postinfection, and GFP levels were measured on day 6 postinfection.  (D)  Similar to the experiment in  (C) , except that T cells were cultured in transwell with EC. Antibody level was at 10 μg/ml. Resting CD4 +  T cells cultured alone or treated with IL-6 served as controls.  (E)  Lack of T cell activation by IL-6 stimulation. Resting CD4 +  T cells were cocultured with IL-6 at 1 ng/ml for 1 day and then infected with a GFP reporter virus. CD25, CD69, and HLA-DR levels were measured on the day of infection (D0) and day 3 (D3) and 6 (D6) postinfection. The cytokine was replenished on days 1, 3, and 5 postinfection, and GFP was measured on day 6 postinfection (the  right  two bars). Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of at least three independent experiments yielding similar results. *Student  t -test;  p

    Journal: AIDS Research and Human Retroviruses

    Article Title: Soluble Factors Secreted by Endothelial Cells Allow for Productive and Latent HIV-1 Infection in Resting CD4+ T Cells

    doi: 10.1089/aid.2016.0058

    Figure Lengend Snippet: IL-6 stimulation increases productive infection in resting CD4 + T cells without T cell activation. (A) Three different cytokines, IL-6, IL-8, and CCL2, were added to resting CD4 + T cells 1 day before infection with a virus expressing GFP. Cytokines were replenished every other day after infection, and GFP-positive rates were measured on day 7 postinfection. (B) IL-6 concentration is positively associated with infection rates. Resting T cells were cultured with various concentrations of IL-6 1 day before infection with a reporter virus. The cytokine was replenished on days 1, 3, and 5 postinfection, and GFP was measured on day 6 postinfection. Resting T cells alone and resting T cells cultured with EC − or EC + were used as controls. (C) EC stimulation is blocked by anti-IL-6 antibody. An anti-IL6 antibody (Ab) was added to EC +/− for approximately an hour before addition of resting T cells in direct contact at three concentrations (10, 5, and 2 μg/ml). After culturing for a day, the cells were infected with a virus expressing GFP. The antibodies were refreshed on days 1 and 3 postinfection, and GFP levels were measured on day 6 postinfection. (D) Similar to the experiment in (C) , except that T cells were cultured in transwell with EC. Antibody level was at 10 μg/ml. Resting CD4 + T cells cultured alone or treated with IL-6 served as controls. (E) Lack of T cell activation by IL-6 stimulation. Resting CD4 + T cells were cocultured with IL-6 at 1 ng/ml for 1 day and then infected with a GFP reporter virus. CD25, CD69, and HLA-DR levels were measured on the day of infection (D0) and day 3 (D3) and 6 (D6) postinfection. The cytokine was replenished on days 1, 3, and 5 postinfection, and GFP was measured on day 6 postinfection (the right two bars). Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of at least three independent experiments yielding similar results. *Student t -test; p

    Article Snippet: ECs treated with or without IFN-γ (EC+/− ) were plated with 100% confluency on the bottom of a 24-well cell culture plate in 0.5 ml of RPMI +10% FBS, and 300 μL of resting CD4+ T cells in the concentration of 1 million per ml was added in Corning Transwell Inserts (24-well inserts, polyester or polycarbonate, pore size of 0.4 μm; Sigma-Aldrich) that rested over the EC cultures.

    Techniques: Infection, Activation Assay, Expressing, Concentration Assay, Cell Culture

    Soluble factors secreted by EC can induce latent HIV infections in resting CD4 +  T cells. Resting CD4 +  T cells were cultured with or without EC in transwells and infected 1 day later with a GFP reporter virus. GFP-negative cells were sorted on day 8 postinfection and treated with or without PMA/I to reactivate latent virus (labeled as induced and uninduced, respectively). GFP was measured 2 days after PMA/I treatment. Samples were taken in duplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results. *Student  t -test;  p

    Journal: AIDS Research and Human Retroviruses

    Article Title: Soluble Factors Secreted by Endothelial Cells Allow for Productive and Latent HIV-1 Infection in Resting CD4+ T Cells

    doi: 10.1089/aid.2016.0058

    Figure Lengend Snippet: Soluble factors secreted by EC can induce latent HIV infections in resting CD4 + T cells. Resting CD4 + T cells were cultured with or without EC in transwells and infected 1 day later with a GFP reporter virus. GFP-negative cells were sorted on day 8 postinfection and treated with or without PMA/I to reactivate latent virus (labeled as induced and uninduced, respectively). GFP was measured 2 days after PMA/I treatment. Samples were taken in duplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results. *Student t -test; p

    Article Snippet: ECs treated with or without IFN-γ (EC+/− ) were plated with 100% confluency on the bottom of a 24-well cell culture plate in 0.5 ml of RPMI +10% FBS, and 300 μL of resting CD4+ T cells in the concentration of 1 million per ml was added in Corning Transwell Inserts (24-well inserts, polyester or polycarbonate, pore size of 0.4 μm; Sigma-Aldrich) that rested over the EC cultures.

    Techniques: Cell Culture, Infection, Labeling

    Resting CD4 +  T cells can be stimulated by EC without direct contact. Resting T cells were cultured alone, in direct contact with EC, or without direct contact, but in transwells with EC. EC +  and EC −  indicate treatment with or without IFN-γ, respectively. Activated T cells (AC) through CD3 and CD28 stimulations were included as a control. All T cells were infected with a HIV reporter virus expressing GFP 1 day after coculture, and the %GFP +  cells were measured on day 7 postinfection. Samples were taken in quadruplicates, and means ± standard errors are plotted. Data shown are the representative of four independent experiments yielding similar results. * p

    Journal: AIDS Research and Human Retroviruses

    Article Title: Soluble Factors Secreted by Endothelial Cells Allow for Productive and Latent HIV-1 Infection in Resting CD4+ T Cells

    doi: 10.1089/aid.2016.0058

    Figure Lengend Snippet: Resting CD4 + T cells can be stimulated by EC without direct contact. Resting T cells were cultured alone, in direct contact with EC, or without direct contact, but in transwells with EC. EC + and EC − indicate treatment with or without IFN-γ, respectively. Activated T cells (AC) through CD3 and CD28 stimulations were included as a control. All T cells were infected with a HIV reporter virus expressing GFP 1 day after coculture, and the %GFP + cells were measured on day 7 postinfection. Samples were taken in quadruplicates, and means ± standard errors are plotted. Data shown are the representative of four independent experiments yielding similar results. * p

    Article Snippet: ECs treated with or without IFN-γ (EC+/− ) were plated with 100% confluency on the bottom of a 24-well cell culture plate in 0.5 ml of RPMI +10% FBS, and 300 μL of resting CD4+ T cells in the concentration of 1 million per ml was added in Corning Transwell Inserts (24-well inserts, polyester or polycarbonate, pore size of 0.4 μm; Sigma-Aldrich) that rested over the EC cultures.

    Techniques: Cell Culture, Infection, Expressing

    Early removal of T cells from EC resulted in reduced productive and latent infections.  (A)  Early removal of resting CD4 +  T cells stimulated by direct contact with EC. Resting T cells were cultured in contact with EC −  or EC + , infected with a GFP reporter virus, and were removed at various time intervals after infection. The %GFP +  cells were measured on day 6 postinfection for all cultures, and the days that the resting CD4 +  T cells were removed postinfection are as indicated. Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results.  (B)  Early removal of resting CD4 +  T cells stimulated by soluble factors with EC. Resting CD4 +  T cells were cultured with EC −  or EC +  in transwells, infected 1 day later with a GFP reporter virus, and removed at the indicated days postinfection. GFP expressions were measured on day 7 after infection. Samples were taken in quadruplicates, and means ± standard errors are plotted. Data shown are the representative of four independent experiments yielding similar results.  (C)  Latent infection in early removal of resting CD4 +  T cells from EC stimulation. Resting CD4 +  T cells were cultured in direct contact with EC, and on various days postinfection, a portion of cells were removed. On day 8 postinfection, GFP-negative cells were sorted and cultured with or without PMA/I (labeled as induced and uninduced, respectively) for 2 days to reactivate latent virus. Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results. *Student  t -test;  p

    Journal: AIDS Research and Human Retroviruses

    Article Title: Soluble Factors Secreted by Endothelial Cells Allow for Productive and Latent HIV-1 Infection in Resting CD4+ T Cells

    doi: 10.1089/aid.2016.0058

    Figure Lengend Snippet: Early removal of T cells from EC resulted in reduced productive and latent infections. (A) Early removal of resting CD4 + T cells stimulated by direct contact with EC. Resting T cells were cultured in contact with EC − or EC + , infected with a GFP reporter virus, and were removed at various time intervals after infection. The %GFP + cells were measured on day 6 postinfection for all cultures, and the days that the resting CD4 + T cells were removed postinfection are as indicated. Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results. (B) Early removal of resting CD4 + T cells stimulated by soluble factors with EC. Resting CD4 + T cells were cultured with EC − or EC + in transwells, infected 1 day later with a GFP reporter virus, and removed at the indicated days postinfection. GFP expressions were measured on day 7 after infection. Samples were taken in quadruplicates, and means ± standard errors are plotted. Data shown are the representative of four independent experiments yielding similar results. (C) Latent infection in early removal of resting CD4 + T cells from EC stimulation. Resting CD4 + T cells were cultured in direct contact with EC, and on various days postinfection, a portion of cells were removed. On day 8 postinfection, GFP-negative cells were sorted and cultured with or without PMA/I (labeled as induced and uninduced, respectively) for 2 days to reactivate latent virus. Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results. *Student t -test; p

    Article Snippet: ECs treated with or without IFN-γ (EC+/− ) were plated with 100% confluency on the bottom of a 24-well cell culture plate in 0.5 ml of RPMI +10% FBS, and 300 μL of resting CD4+ T cells in the concentration of 1 million per ml was added in Corning Transwell Inserts (24-well inserts, polyester or polycarbonate, pore size of 0.4 μm; Sigma-Aldrich) that rested over the EC cultures.

    Techniques: Cell Culture, Infection, Labeling

    Levels of IL-6, IL-8, and CCL2 in cultures of resting CD4 +  T cells stimulated by EC. Resting CD4 +  T cells were cultured alone, with MCF7, or with EC + /EC −  for 6 days, and the supernatants were collected and analyzed for 30 cytokines.  (A)  IL-6 levels in resting T cells cultured alone or stimulated by EC. EC cultured alone were included as well.  (B)  IL-8 levels in resting T cells cultured alone, with MCF7 or stimulated by EC.  (C)  CCL2 levels in resting CD4 +  T cells cultured alone, with MCF7 or stimulated by EC. Samples were averaged from two different donors each in duplicates, and means ± standard errors are plotted.

    Journal: AIDS Research and Human Retroviruses

    Article Title: Soluble Factors Secreted by Endothelial Cells Allow for Productive and Latent HIV-1 Infection in Resting CD4+ T Cells

    doi: 10.1089/aid.2016.0058

    Figure Lengend Snippet: Levels of IL-6, IL-8, and CCL2 in cultures of resting CD4 + T cells stimulated by EC. Resting CD4 + T cells were cultured alone, with MCF7, or with EC + /EC − for 6 days, and the supernatants were collected and analyzed for 30 cytokines. (A) IL-6 levels in resting T cells cultured alone or stimulated by EC. EC cultured alone were included as well. (B) IL-8 levels in resting T cells cultured alone, with MCF7 or stimulated by EC. (C) CCL2 levels in resting CD4 + T cells cultured alone, with MCF7 or stimulated by EC. Samples were averaged from two different donors each in duplicates, and means ± standard errors are plotted.

    Article Snippet: ECs treated with or without IFN-γ (EC+/− ) were plated with 100% confluency on the bottom of a 24-well cell culture plate in 0.5 ml of RPMI +10% FBS, and 300 μL of resting CD4+ T cells in the concentration of 1 million per ml was added in Corning Transwell Inserts (24-well inserts, polyester or polycarbonate, pore size of 0.4 μm; Sigma-Aldrich) that rested over the EC cultures.

    Techniques: Cell Culture

    JNK, Murr1, and Glut1 expression in resting T cells does not change after stimulation by EC.  (A)  Resting T cells were cultured alone, with EC +/− , or with PMA and Ionomycin. p-JNK intracellular staining was done 4 days later for PMA/I-activated cells, and 7 days later for resting T cells cultured alone or with EC −  and EC + . Samples were taken in duplicates, and means ± standard deviations are plotted. Data shown are the representative of two independent experiments yielding similar results.  (B)  In a similar way, resting CD4 +  T cells were cultured alone, activated with PMA/I, or cocultured with EC −  or EC +  and analyzed for JNK expression through western blot analysis.  (C)  Resting CD4 +  T cells were cultured alone or with EC −  or EC +  and infected with a GFP reporter virus the next day. Intracellular staining was done on day 7 postinfection to measure p-JNK expression. GFP +  cells were productively infected, and GFP −  cells were not productively infected. Samples were taken in duplicates, and means ± standard deviations are plotted.  (D)  Resting T cells cultured alone or with EC +/−  or activated with PMA/I. Murr1 expression was determined by intracellular staining 7 days after infection. Samples were taken in duplicates, and means ± standard deviations are plotted. Data shown are the representative of two independent experiments yielding similar results.  (E)  Resting CD4 +  T cells were cultured alone, activated by PHA, or stimulated by EC +/− . Cells were infected with a GFP reporter virus. GFP expression and GLUT1 staining were performed 3 days after infection for PHA-activated cells and 7 days after infection for resting cells that were unstimulated or stimulated by EC. Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results. *Student t-test;  p

    Journal: AIDS Research and Human Retroviruses

    Article Title: Soluble Factors Secreted by Endothelial Cells Allow for Productive and Latent HIV-1 Infection in Resting CD4+ T Cells

    doi: 10.1089/aid.2016.0058

    Figure Lengend Snippet: JNK, Murr1, and Glut1 expression in resting T cells does not change after stimulation by EC. (A) Resting T cells were cultured alone, with EC +/− , or with PMA and Ionomycin. p-JNK intracellular staining was done 4 days later for PMA/I-activated cells, and 7 days later for resting T cells cultured alone or with EC − and EC + . Samples were taken in duplicates, and means ± standard deviations are plotted. Data shown are the representative of two independent experiments yielding similar results. (B) In a similar way, resting CD4 + T cells were cultured alone, activated with PMA/I, or cocultured with EC − or EC + and analyzed for JNK expression through western blot analysis. (C) Resting CD4 + T cells were cultured alone or with EC − or EC + and infected with a GFP reporter virus the next day. Intracellular staining was done on day 7 postinfection to measure p-JNK expression. GFP + cells were productively infected, and GFP − cells were not productively infected. Samples were taken in duplicates, and means ± standard deviations are plotted. (D) Resting T cells cultured alone or with EC +/− or activated with PMA/I. Murr1 expression was determined by intracellular staining 7 days after infection. Samples were taken in duplicates, and means ± standard deviations are plotted. Data shown are the representative of two independent experiments yielding similar results. (E) Resting CD4 + T cells were cultured alone, activated by PHA, or stimulated by EC +/− . Cells were infected with a GFP reporter virus. GFP expression and GLUT1 staining were performed 3 days after infection for PHA-activated cells and 7 days after infection for resting cells that were unstimulated or stimulated by EC. Samples were taken in triplicates, and means ± standard errors are plotted. Data shown are the representative of three independent experiments yielding similar results. *Student t-test; p

    Article Snippet: ECs treated with or without IFN-γ (EC+/− ) were plated with 100% confluency on the bottom of a 24-well cell culture plate in 0.5 ml of RPMI +10% FBS, and 300 μL of resting CD4+ T cells in the concentration of 1 million per ml was added in Corning Transwell Inserts (24-well inserts, polyester or polycarbonate, pore size of 0.4 μm; Sigma-Aldrich) that rested over the EC cultures.

    Techniques: Expressing, Cell Culture, Staining, Western Blot, Infection

    Overexpression of Fbxl8 recedes G1-S phase transition of cell cycle through degradation of cyclin D3. a Schematic model of experiment. NIH3T3 cells were transfected with MigR1 IRES-GFP, MigR1Fbxl8 IRES-GFP or MigR1Fbxl8ΔF IRES-GFP, and arrested at G0/G1 phase by serum starvation for 36 h. GFP positive cells and negative cells were FACS sorted and S phase entry was assessed by BrdU incorporation (30 min). b Western analysis of lysates from sorted GFP positive NIH3T3 cells expressing GFP, Fbxl8, or Fbxl8ΔF for cyclin D3, cyclin D1, Flag-Fbxl8 and βactin. The numbers indicate quantifications of cyclin D3 and cyclin D1 normalized by βactin. c GFP and BrdU double-positive cells were analyzed 9, 12, 15, 18, 21 h after release from G0/G1 phase by re-splitting cells in DMEM with 10%FBS. Quantification of GFP and BrdU double-positive cells; mean ± SD, * p

    Journal: Oncogene

    Article Title: Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3

    doi: 10.1038/s41388-020-01532-4

    Figure Lengend Snippet: Overexpression of Fbxl8 recedes G1-S phase transition of cell cycle through degradation of cyclin D3. a Schematic model of experiment. NIH3T3 cells were transfected with MigR1 IRES-GFP, MigR1Fbxl8 IRES-GFP or MigR1Fbxl8ΔF IRES-GFP, and arrested at G0/G1 phase by serum starvation for 36 h. GFP positive cells and negative cells were FACS sorted and S phase entry was assessed by BrdU incorporation (30 min). b Western analysis of lysates from sorted GFP positive NIH3T3 cells expressing GFP, Fbxl8, or Fbxl8ΔF for cyclin D3, cyclin D1, Flag-Fbxl8 and βactin. The numbers indicate quantifications of cyclin D3 and cyclin D1 normalized by βactin. c GFP and BrdU double-positive cells were analyzed 9, 12, 15, 18, 21 h after release from G0/G1 phase by re-splitting cells in DMEM with 10%FBS. Quantification of GFP and BrdU double-positive cells; mean ± SD, * p

    Article Snippet: Cell culture, transfection, infection, cell cycle analyses and CRISPR-Cas9 knockoutNIH3T3 and 293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 units/ml of penicillin, and 100 mg/ml of streptomycin (Corning).

    Techniques: Over Expression, Sublimation, Transfection, FACS, BrdU Incorporation Assay, Western Blot, Expressing

    Silence of FOXO1 mitigates the inhibitory effects of miR‐5188 knockdown on glioma cell proliferation, invasion and migration. MTT assays (A), EdU incorporation assays (B), flow cytometry assays (C), transwell assays (D) and Boyden assays (E) were performed in U251 or U87 cells treated with control sequence, FOXO1 siRNA or miR‐5188 inhibitor, as indicated. Scale bars, 100 μm. F, Western blotting analyses were applied to measure the protein levels of FOXO1, P‐PI3K, PI3K, P‐AKT, AKT, CCND1, CDK4, c‐JUN, N‐cadherin and vimentin in U251 or U87 cells treated with control sequence, FOXO1 siRNA or miR‐5188 inhibitor, as indicated. G, Western blotting analysis analyses were applied to measure the protein levels of FOXO1, P‐PI3K, PI3K, P‐AKT, AKT, CCND1, CDK4, c‐JUN, N‐cadherin and vimentin protein levels in U87 and U251 cells transfected with control sequence or FOXO1 siRNA. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: miR‐5188 augments glioma growth, migration and invasion through an SP1‐modulated FOXO1‐PI3K/AKT‐c‐JUN‐positive feedback circuit, et al. miR‐5188 augments glioma growth, migration and invasion through an SP1‐modulated FOXO1‐PI3K/AKT‐c‐JUN‐positive feedback circuit

    doi: 10.1111/jcmm.15794

    Figure Lengend Snippet: Silence of FOXO1 mitigates the inhibitory effects of miR‐5188 knockdown on glioma cell proliferation, invasion and migration. MTT assays (A), EdU incorporation assays (B), flow cytometry assays (C), transwell assays (D) and Boyden assays (E) were performed in U251 or U87 cells treated with control sequence, FOXO1 siRNA or miR‐5188 inhibitor, as indicated. Scale bars, 100 μm. F, Western blotting analyses were applied to measure the protein levels of FOXO1, P‐PI3K, PI3K, P‐AKT, AKT, CCND1, CDK4, c‐JUN, N‐cadherin and vimentin in U251 or U87 cells treated with control sequence, FOXO1 siRNA or miR‐5188 inhibitor, as indicated. G, Western blotting analysis analyses were applied to measure the protein levels of FOXO1, P‐PI3K, PI3K, P‐AKT, AKT, CCND1, CDK4, c‐JUN, N‐cadherin and vimentin protein levels in U87 and U251 cells transfected with control sequence or FOXO1 siRNA. * P

    Article Snippet: For the cell migration assay, a total of 1 × 104 cells in 100 μL DMEM medium without FBS were seeded on a Transwell apparatus (Costar, MA).

    Techniques: Migration, MTT Assay, Flow Cytometry, Sequencing, Western Blot, Transfection

    miR‐5188 is elevated in glioma tissues and promotes glioma cell proliferation, migration and invasion in vitro by up‐regulating PI3K/AKT activity. A, Increased levels of miR‐5188, as assessed using real‐time quantitative PCR (RT‐qPCR), were positively correlated with the pathology classification status. An unpaired t test was applied for assessing the results of this assay in normal tissues (n = 17), grade II glioma (n = 12), grade III glioma (n = 13) and grade IV glioma (n = 15). An MTT assay (B), colony formation assay (C), cell cycle analysis (D) and EdU incorporation assay (E) were applied to investigate the impact of miR‐5188 on cell proliferation in U87 and U251. Scale bars, 100 μm. F, A transwell assay and Boyden chamber assays were applied to elucidate the impact of miR‐5188 on cell migration and invasion in U87 and U251. Scale bars, 100 μm. G, Western blotting analyses were applied to assess P‐PI3K, PI3K, P‐AKT, AKT, CDK4, CCND1, c‐JUN, N‐cadherin and vimentin protein levels in U87 and U251 cells treated with miR‐5188 inhibitor or mimics. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: miR‐5188 augments glioma growth, migration and invasion through an SP1‐modulated FOXO1‐PI3K/AKT‐c‐JUN‐positive feedback circuit, et al. miR‐5188 augments glioma growth, migration and invasion through an SP1‐modulated FOXO1‐PI3K/AKT‐c‐JUN‐positive feedback circuit

    doi: 10.1111/jcmm.15794

    Figure Lengend Snippet: miR‐5188 is elevated in glioma tissues and promotes glioma cell proliferation, migration and invasion in vitro by up‐regulating PI3K/AKT activity. A, Increased levels of miR‐5188, as assessed using real‐time quantitative PCR (RT‐qPCR), were positively correlated with the pathology classification status. An unpaired t test was applied for assessing the results of this assay in normal tissues (n = 17), grade II glioma (n = 12), grade III glioma (n = 13) and grade IV glioma (n = 15). An MTT assay (B), colony formation assay (C), cell cycle analysis (D) and EdU incorporation assay (E) were applied to investigate the impact of miR‐5188 on cell proliferation in U87 and U251. Scale bars, 100 μm. F, A transwell assay and Boyden chamber assays were applied to elucidate the impact of miR‐5188 on cell migration and invasion in U87 and U251. Scale bars, 100 μm. G, Western blotting analyses were applied to assess P‐PI3K, PI3K, P‐AKT, AKT, CDK4, CCND1, c‐JUN, N‐cadherin and vimentin protein levels in U87 and U251 cells treated with miR‐5188 inhibitor or mimics. * P

    Article Snippet: For the cell migration assay, a total of 1 × 104 cells in 100 μL DMEM medium without FBS were seeded on a Transwell apparatus (Costar, MA).

    Techniques: Migration, In Vitro, Activity Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, MTT Assay, Colony Assay, Cell Cycle Assay, Transwell Assay, Western Blot