Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Impaired Adipose Anabolism in Pancreatic Cancer Cachexia Is Reversed by HuR Inhibition

doi: 10.1002/jcsm.70253

Figure Lengend Snippet: Adipocytes suppress lipogenesis in response to pancreatic ductal adenocarcinoma‐conditioned media. (A) Representative bright‐field images of differentiated 3T3‐L1 cells treated with control media (top) or KPC CM (bottom) for 6 days. Scale bars represent 70 μm. (B) Quantification of Oil Red O staining. N = 4 wells per condition. (C–E) 3T3‐L1 cells were fully differentiated and then treated with control, KPC‐conditioned or isoproterenol (10 μM) media for 1, 3 and 24 h in the absence of FBS and insulin. N = 3 wells per timepoint/condition. (C) Media glycerol levels, normalized by hours of collection. Conditioned media and control media alone did not contain glycerol above background (assay buffer only) levels. (D) Serine 563 phosphorylated HSL protein levels normalized to total HSL protein levels. (E) Serine 660 phosphorylated HSL protein levels normalized to total HSL protein levels. (F) mRNA expression of lipolysis and lipogenesis genes. N = 3 wells per condition, normalized to 18S expression. Panels (B) and (F) tested with t ‐test. Panels (C–E) tested with two‐way ANOVA with Tukey correction for multiple comparisons. p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: 3T3‐L1 adipocytes (ATCC CL‐173) were purchased from ATCC and stored in liquid nitrogen until use.

Techniques: Control, Staining, Expressing

Journal: Nutrients

Article Title: Severity of Hypoxia-Induced Effects on 3T3-L1 Adipocyte Secretory Function Is Attenuated Dose-Dependently by Individual Short-Chain Fatty Acids

doi: 10.3390/nu18060942

Figure Lengend Snippet: Secreted adipokine levels from 3T3-L1 adipocytes in response to the dose-dependent addition of individual SCFAs (Ace, Pro and But at 0.25 mM, 0.5 mM and 1 mM) in hypoxic environmental conditions (10 µM CoCl 2 ; black bars). Normoxic control cultures (without CoCl 2 ) served as negative controls (gray bars). Bars represent mean values ± SEM ( n = 8–9/experimental group). Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison test. Bars not sharing a lower-case letter differ ( p < 0.05).

Article Snippet: 3T3-L1 murine pre-adipocytes (CL-173; American Type Culture Collection, Manassas, VA, USA) were cultured and maintained according to the manufacturer’s instructions in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Logan, UT, USA) supplemented with 4 mM L-glutamine, 4500 mg/L glucose, 10% ( v / v ) low endotoxin sterile-filtered fetal bovine serum (FBS; Millipore-Sigma, Oakville, ON, Canada) and 1% ( v / v ) penicillin-streptomycin (Fisher Scientific, Mississauga, ON, Canada), as described previously [ , , ].

Techniques: Control, Comparison

Journal: Nutrients

Article Title: Severity of Hypoxia-Induced Effects on 3T3-L1 Adipocyte Secretory Function Is Attenuated Dose-Dependently by Individual Short-Chain Fatty Acids

doi: 10.3390/nu18060942

Figure Lengend Snippet: Secreted adipokine levels (i.e., cytokines and chemokines) from 3T3-L1 adipocytes in response to the dose-dependent [0.25 mM, 0.5 mM and 1 mM] addition of individual SCFAs (Ace, Pro and But) in combined inflammatory and hypoxic environmental conditions (+LPS/CoCl 2 ). Bars represent mean values ± SEM ( n = 8–9/experimental group). Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison test. Bars not sharing a lower-case letter differ ( p < 0.05).

Article Snippet: 3T3-L1 murine pre-adipocytes (CL-173; American Type Culture Collection, Manassas, VA, USA) were cultured and maintained according to the manufacturer’s instructions in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Logan, UT, USA) supplemented with 4 mM L-glutamine, 4500 mg/L glucose, 10% ( v / v ) low endotoxin sterile-filtered fetal bovine serum (FBS; Millipore-Sigma, Oakville, ON, Canada) and 1% ( v / v ) penicillin-streptomycin (Fisher Scientific, Mississauga, ON, Canada), as described previously [ , , ].

Techniques: Comparison

Journal: Nutrients

Article Title: Severity of Hypoxia-Induced Effects on 3T3-L1 Adipocyte Secretory Function Is Attenuated Dose-Dependently by Individual Short-Chain Fatty Acids

doi: 10.3390/nu18060942

Figure Lengend Snippet: Secreted adipokine levels (i.e., resistin, leptin and adiponectin) from 3T3-L1 adipocytes in response to the dose-dependent [0.25 mM, 0.5 mM and 1 mM] addition of individual SCFAs (Ace, Pro and But) in combined inflammatory and hypoxic environmental conditions (+LPS/CoCl 2 ). Bars represent mean values ± SEM ( n = 8–9/experimental group). Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison test. Bars not sharing a lower-case letter differ ( p < 0.05).

Article Snippet: 3T3-L1 murine pre-adipocytes (CL-173; American Type Culture Collection, Manassas, VA, USA) were cultured and maintained according to the manufacturer’s instructions in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Logan, UT, USA) supplemented with 4 mM L-glutamine, 4500 mg/L glucose, 10% ( v / v ) low endotoxin sterile-filtered fetal bovine serum (FBS; Millipore-Sigma, Oakville, ON, Canada) and 1% ( v / v ) penicillin-streptomycin (Fisher Scientific, Mississauga, ON, Canada), as described previously [ , , ].

Techniques: Comparison

Journal: Nutrients

Article Title: Severity of Hypoxia-Induced Effects on 3T3-L1 Adipocyte Secretory Function Is Attenuated Dose-Dependently by Individual Short-Chain Fatty Acids

doi: 10.3390/nu18060942

Figure Lengend Snippet: 3T3-L1 adipocyte intracellular protein expression of the ratio of phosphorylated to total NFκB p65 and STAT3 in response to the dose-dependent [0.25 mM, 0.5 mM and 1 mM] addition of individual SCFAs (Ace, Pro and But) in combined inflammatory and hypoxic environmental conditions (+LPS/CoCl 2 ). Bars represent mean values ± SEM ( n = 8–9/experimental group). Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison test. Bars not sharing a lower-case letter differ ( p < 0.05).

Article Snippet: 3T3-L1 murine pre-adipocytes (CL-173; American Type Culture Collection, Manassas, VA, USA) were cultured and maintained according to the manufacturer’s instructions in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Logan, UT, USA) supplemented with 4 mM L-glutamine, 4500 mg/L glucose, 10% ( v / v ) low endotoxin sterile-filtered fetal bovine serum (FBS; Millipore-Sigma, Oakville, ON, Canada) and 1% ( v / v ) penicillin-streptomycin (Fisher Scientific, Mississauga, ON, Canada), as described previously [ , , ].

Techniques: Expressing, Comparison

Journal: Nutrients

Article Title: Severity of Hypoxia-Induced Effects on 3T3-L1 Adipocyte Secretory Function Is Attenuated Dose-Dependently by Individual Short-Chain Fatty Acids

doi: 10.3390/nu18060942

Figure Lengend Snippet: 2-dexoy glucose uptake by 3T3-L1 adipocytes treated with 1 mM Ace, Pro or But under combined inflammatory and hypoxic environmental conditions (+LPS/CoCl 2 ) in basal (i.e., non-insulin stimulated: phosphate-buffered saline for 20 min followed by 10 mM 2-deoxy glucose for 20 min at 37 °C) and insulin-stimulated (1 µM insulin for 20 min followed by 10 mM 2-deoxy glucose for 20 min at 37 °C) conditions ( n = 8–9/treatment group: CON, Ace, Pro and But in either basal or insulin-stimulated condition). Bars represent mean values ± SEM. Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison test. Bars not sharing a lower-case letter differ ( p < 0.05).

Article Snippet: 3T3-L1 murine pre-adipocytes (CL-173; American Type Culture Collection, Manassas, VA, USA) were cultured and maintained according to the manufacturer’s instructions in Dulbecco’s modified Eagle’s medium (DMEM; HyClone, Logan, UT, USA) supplemented with 4 mM L-glutamine, 4500 mg/L glucose, 10% ( v / v ) low endotoxin sterile-filtered fetal bovine serum (FBS; Millipore-Sigma, Oakville, ON, Canada) and 1% ( v / v ) penicillin-streptomycin (Fisher Scientific, Mississauga, ON, Canada), as described previously [ , , ].

Techniques: Saline, Comparison