fbs  (Gemini Bio)

Journal: Arthritis Research & Therapy

Article Title: Arhalofenate acid inhibits monosodium urate crystal-induced inflammatory responses through activation of AMP-activated protein kinase (AMPK) signaling

doi: 10.1186/s13075-018-1699-4

Figure Lengend Snippet: Arhalofenate acid activated AMPK downstream targets involved in regulation of mitochondrial function and maintained mitochondrial cristae area. BMDMs were treated with arhalofenate acid (100 μM) for 1 h before stimulation with monosodium urate (MSU) crystals (0.2 mg/mL) for 1 h or 18 h in RPMI containing 1% FBS. Cell lysates prepared from 18-h treatment samples were subjected to Western blot analysis of phosphorylation (p) and expression of AMP-activated protein kinase (AMPK)α, expression of sirtuin 1 (SIRT1), peroxisome proliferator-activated receptor γ co-activator 1α (PGC-1α), and mitochondrial transcription factor A (TFAM), and expression of thioredoxin (TRX)1, TRX2, and thioredoxin-interacting protein (TXNIP) ( a ). Cells from 1-h treatment were then stained with MitoSOX Red and MitoTracker Green and visualized by fluorescence microscopy ( b , magnification 20×). TEM analysis was performed to examine mitochondrial cristae (the folds of inner mitochondrial membrane indicated by white arrows in c ), and the cristae volume density are presented ( d ). Data in a and b are representative of three individual experiments. Data in c are representative of 30 cells examined for each condition. Data in d are the mean ± SEM of 30 cells. The p values represent comparisons between none and MSU crystals alone, or between MSU crystals alone and MSU crystals plus arhalofenate acid

Article Snippet: Briefly, bone marrow cells were cultured in complete RPMI media containing 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 μg/mL) in the presence of macrophage colony-stimulating factor (M-CSF; 20 ng/mL; Gemini Bio-products, West Sacramento, CA).

Techniques: Western Blot, Expressing, Pyrolysis Gas Chromatography, Staining, Fluorescence, Microscopy, Transmission Electron Microscopy

Journal: Arthritis Research & Therapy

Article Title: Arhalofenate acid inhibits monosodium urate crystal-induced inflammatory responses through activation of AMP-activated protein kinase (AMPK) signaling

doi: 10.1186/s13075-018-1699-4

Figure Lengend Snippet: Arhalofenate acid prevented prolonged accumulation of p62 by promoting autophagy flux in response to MSU crystals. BMDMs were treated with arhalofenate acid (100 μM) for 1 h before stimulated with monosodium urate (MSU) crystals (0.2 mg/mL) for 2 h in the presence or absence of bafilomycin (Baf; 100 nM) and for 6 h in RPMI containing 1% FBS. Cell lysates were subjected to Western blot analysis of LC3 and p62 ( a ). TEM was performed to examine autophagosomes (indicated by black arrows in b ), and the numbers of autophagosomes containing electron dense material per μm 2 are presented ( c ). Data in a and b are representative of three individual experiments. Data in c are the mean ± SEM of 30 cells. The p values represent comparisons between none and MSU crystals alone, or between MSU crystals alone and MSU crystals plus arhalofenate acid

Article Snippet: Briefly, bone marrow cells were cultured in complete RPMI media containing 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 μg/mL) in the presence of macrophage colony-stimulating factor (M-CSF; 20 ng/mL; Gemini Bio-products, West Sacramento, CA).

Techniques: Western Blot, Transmission Electron Microscopy

Journal: Arthritis Research & Therapy

Article Title: Arhalofenate acid inhibits monosodium urate crystal-induced inflammatory responses through activation of AMP-activated protein kinase (AMPK) signaling

doi: 10.1186/s13075-018-1699-4

Figure Lengend Snippet: Arhalofenate acid attenuates MSU crystal-induced IL-1β release by inhibiting NLRP3 inflammasome activation in BMDMs in vitro. BMDMs were pretreated with arhalofenate acid at a concentration of 100 μM for 1 h before being stimulated with monosodium urate (MSU) crystals (0.2 mg/mL) in RPMI containing 1% FBS for 18 h. The conditioned media was used for ELISA for interleukin (IL)-1β ( a ), and the cell lysates were subjected to Western blot analysis ( b ) for expression of NLRP3, pro-caspase 1, and cleaved caspase 1 (p10). Data in a are the mean ± SD of three individual experiments, and p values represent comparisons between none and MSU crystals alone, or between MSU crystal alone and MSU crystals plus arhalofenate acid. Data in b are representative of three individual experiments

Article Snippet: Briefly, bone marrow cells were cultured in complete RPMI media containing 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 μg/mL) in the presence of macrophage colony-stimulating factor (M-CSF; 20 ng/mL; Gemini Bio-products, West Sacramento, CA).

Techniques: Activation Assay, In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Arthritis Research & Therapy

Article Title: Arhalofenate acid inhibits monosodium urate crystal-induced inflammatory responses through activation of AMP-activated protein kinase (AMPK) signaling

doi: 10.1186/s13075-018-1699-4

Figure Lengend Snippet: Arhalofenate acid inhibited MSU crystal-induced IL-1β via AMPK in BMDMs in vitro. BMDMs were treated with arhalofenate acid at 100 μM for 1 h before being stimulated with monosodium urate (MSU) crystals (0.2 mg/mL) for 18 h in RPMI containing 1% FBS. Cell lysates were subjected to Western blot analysis for phosphorylation (p) and expression of AMP-activated protein kinase (AMPK)α ( a ). The conditioned medium was used for ELISA analysis of interleukin (IL)-1β release ( b ). Data in a are representative of three individual experiments. Data in b are the mean ± SD of three individual experiments. The p values in b represent comparisons between none and MSU crystals alone in the presence or absence of arhalofenate acid in either wild-type (WT) or AMPKα1 knockout (KO) BMDMs. ns not significant

Article Snippet: Briefly, bone marrow cells were cultured in complete RPMI media containing 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 μg/mL) in the presence of macrophage colony-stimulating factor (M-CSF; 20 ng/mL; Gemini Bio-products, West Sacramento, CA).

Techniques: In Vitro, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Knock-Out

Journal: Arthritis Research & Therapy

Article Title: Arhalofenate acid inhibits monosodium urate crystal-induced inflammatory responses through activation of AMP-activated protein kinase (AMPK) signaling

doi: 10.1186/s13075-018-1699-4

Figure Lengend Snippet: Arhalofenate acid induced phosphorylation of AMPKα and expression of SIRT1 in BMDMs in vitro. BMDMs prepared from wild-type (WT) and AMPKα1 knockout (KO) mice were pretreated with arhalofenate acid at the concentrations indicated for 18 h ( a ) or at 100 μM for 1 h ( b ) in RPMI containing 1% FBS. Cell lysates were subjected to Western blot analysis for phosphorylation (p) and expression of AMP-activated protein kinase (AMPK)α and sirtuin 1 (SIRT1). Data in both a and b are representative of three individual experiments

Article Snippet: Briefly, bone marrow cells were cultured in complete RPMI media containing 10% fetal bovine serum (FBS), penicillin (100 U/mL), and streptomycin (100 μg/mL) in the presence of macrophage colony-stimulating factor (M-CSF; 20 ng/mL; Gemini Bio-products, West Sacramento, CA).

Techniques: Expressing, In Vitro, Knock-Out, Mouse Assay, Western Blot

Journal: Cell

Article Title: p53 Represses the Mevalonate Pathway to Mediate Tumor Suppression

doi: 10.1016/j.cell.2018.11.011

Figure Lengend Snippet: p53 Inhibits SREBP-2 Maturation under Low-Sterol Conditions (A) HCT116 cells were cultured for 24 hr in medium plus fetal bovine serum (FBS; gray bars), delipidated FBS (DL-FBS; black bars), or delipidated FBS plus 25-hydroxycholesterol (DL-FBS+25-HC; white bars). Expression of HMGCR , HMGCS1 , and SREBF-2 was assessed by qRT-PCR (n = 3; *p

Article Snippet: For analysis of cell cycle changes during sterol starvation, cells were rinsed once with serum-free medium and then placed in medium with 10% delipidated FBS (Gemini Bio-products, catalog #900–123).

Techniques: Cell Culture, Expressing, Quantitative RT-PCR

Journal:

Article Title: At diabetes-like concentration, glucose down-regulates the placental serotonin transport system in a cell-cycle-dependent manner

doi: 10.1111/j.1471-4159.2007.04469.x

Figure Lengend Snippet: Characterization of SERT in JAR cells with glucose at different concentrations. (a) JAR cells were cultured in RPMI supplemented with 10% FBS with/without insulin and D-glucose at different concentrations, 5.5, 10, 15, 25 mmol/L. The [ 3 H]-5HT uptake rate

Article Snippet: Also, these data demonstrate that there is a 60% increase in overall DNA synthesis of JAR cells if they enter the cell cycle in media supplemented with 25 mmol/L D-glucose and insulin-free FBS ( ).

Techniques: Cell Culture

Journal:

Article Title: At diabetes-like concentration, glucose down-regulates the placental serotonin transport system in a cell-cycle-dependent manner

doi: 10.1111/j.1471-4159.2007.04469.x

Figure Lengend Snippet: Concentration-dependent effect of exogenous D-glucose on relative SERT mRNA in JAR cells. JAR cells were cultured in RPMI supplemented with 10% FBS without insulin and D-glucose at different concentrations, 5.5, 10, 15, 25 mmol/L (lanes 2–5).

Article Snippet: Also, these data demonstrate that there is a 60% increase in overall DNA synthesis of JAR cells if they enter the cell cycle in media supplemented with 25 mmol/L D-glucose and insulin-free FBS ( ).

Techniques: Concentration Assay, Cell Culture

Journal: Stem Cell Research & Therapy

Article Title: Influence of cholesterol/caveolin-1/caveolae homeostasis on membrane properties and substrate adhesion characteristics of adult human mesenchymal stem cells

doi: 10.1186/s13287-018-0830-4

Figure Lengend Snippet: Effects of methyl-β-cyclodextrin (MβCD) and cholesterol (chol)-MβCD treatments on membrane cholesterol level and cell viability of MSCs. a Cholesterol level of MSCs after treatment with MβCD for 40 min at the concentrations indicated. Membrane cholesterol levels were measured and compared (untreated cells = 100%). b Cholesterol level of MSCs after treatment with 10 mM MβCD or 100 μM chol-MβCD for the time indicated. c Time course of cholesterol level of MSCs after initial treatment with 10 mM MβCD for 60 min. After treatment, cells were washed with PBS and incubated in fresh PM containing CS-FBS for the time indicated. d Viability of cells, treated as described in Fig. 1b, evaluated by MTS assay. Results are presented as mean ± SD; n ≥ 3 independent experiments per cell line per time point. The results showed the effectiveness of MβCD and chol-MβCD in altering MSC membrane cholesterol level without compromising cell viability. * p

Article Snippet: After MβCD pretreatment, cholesterol-depleted cells were cultured in fresh medium containing charcoal stripped FBS (CS-FBS; Gemini Bio-Products, West Sacramento, CA, USA).

Techniques: Incubation, MTS Assay