Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Adipokines and Vascular Endothelial Growth Factor in Normal Human Breast Tissue in Vivo – Correlations and Attenuation by Dietary Flaxseed

doi: 10.1007/s10911-016-9352-9

Figure Lengend Snippet: Inhibition of VEGF decreased leptin secretion in co-culture of MCF7 and adipocytes. Co-culture of ER+ breast cancer cells, MCF7, and freshly isolated human adipocytes, were exposed to; a . Anti-VEGF antibody at 0.1 μg/ml and 1 μg/ml. Extracellular levels of leptin were analyzed after 48 h. *, p < 0.05 and ****, p < 0.0001 compared with controls, n = 5–6 in each group. b . Anti-leptin antibody at 0.1 μg/ml and 1 μg/ml. Extracellular levels of VEGF were analyzed after 48 h. n = 5–6 in each group. c . Anti-VEGF antibody at 0.1 μg/ml and 1 μg/ml in the presence of estradiol (10 −9 M). Extracellular levels of leptin were analyzed after 48 h. ***, p < 0.001 and ****, p < 0.0001 compared with controls, n = 5–6 in each group. d . Anti-leptin antibody at 0.1 μg/ml and 1 μg/ml in presence of estradiol (10 −9 M). Extracellular levels of VEGF were analyzed after 48 h. n = 5–6 in each group

Article Snippet: MCF7 cells (HTB-22; Human breast adenocarcinoma, estrogen receptor (ER) and progesterone receptor positive), obtained from the American type culture collection (ATCC; Manassas, Va, USA), were maintained in DMEM without phenol red (Invitrogen, Carlsbad, CA, USA) supplemented with 10 % fetal bovine serum (FBS) (Invitrogen), 2 mM glutamine (Invitrogen) 50 IU/ml penicillin-G and 50 μg/ml streptomycin (Invitrogen), at 37 °C in a humidified atmosphere of 5 % CO 2 .

Techniques: Inhibition, Co-Culture Assay, Isolation

Journal: Journal of Mammary Gland Biology and Neoplasia

Article Title: Adipokines and Vascular Endothelial Growth Factor in Normal Human Breast Tissue in Vivo – Correlations and Attenuation by Dietary Flaxseed

doi: 10.1007/s10911-016-9352-9

Figure Lengend Snippet: Co-culture with adipocytes enhanced the effects of estradiol on leptin and VEGF secretion. A. MCF7 cells, freshly isolated human adipocytes or co-culture of MCF7 and adipocytes, were cultured with or without estradiol (10 −9 M) for 48 h. Extracellular levels of VEGF were measured in the conditioned medium with ELISA. ***, p < 0.001 and ****, p < 0.0001, n = 6. B. MCF7 cells alone or co-cultured with adipocytes were treated with estradiol (10 −9 M) or estradiol (10 −9 M) and fulvestrand (10 −6 M) for 48 h. Extracellular levels of VEGF were measured with ELISA in the conditioned medium. ***, p < 0.001 and ****, p < 0.0001, n = 6. C. MCF7 cells, adipocytes or co-culture of MCF7 and adipocytes, were cultured with or without estradiol (10 −9 M) for 48 h. Extracellular levels of leptin were measured with ELISA in the conditioned medium. ***, p < 0.001, n = 6. D. MCF7 cells alone or co-cultured with adipocytes were treated with estradiol (10 −9 M) or estradiol (10 −9 M) and fulvestrant (10 −6 M) for 48 h. Extracellular levels of leptin were measured with ELISA in the conditioned medium. *, p < 0.05 and ***, p < 0.001, n = 6. E. MCF7 cells, adipocytes or co-culture of MCF7 and adipocytes, were cultured with or without estradiol (10 −9 M) for 48 h. Extracellular levels of adiponctin were measured with ELISA in the conditioned medium. n = 6 in each group

Article Snippet: MCF7 cells (HTB-22; Human breast adenocarcinoma, estrogen receptor (ER) and progesterone receptor positive), obtained from the American type culture collection (ATCC; Manassas, Va, USA), were maintained in DMEM without phenol red (Invitrogen, Carlsbad, CA, USA) supplemented with 10 % fetal bovine serum (FBS) (Invitrogen), 2 mM glutamine (Invitrogen) 50 IU/ml penicillin-G and 50 μg/ml streptomycin (Invitrogen), at 37 °C in a humidified atmosphere of 5 % CO 2 .

Techniques: Co-Culture Assay, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Serial Examination of an Inducible and Reversible Dilated Cardiomyopathy in Individual Adult Drosophila

doi: 10.1371/journal.pone.0007132

Figure Lengend Snippet: Cardiac parameters in w 1118 , Oregon-R , and the cardiac-specific, inducible driver line.

Article Snippet: Applied Biosystems Taqman Gene expression assays were used to perform quantitative (real time) RT-PCR (human delta-sarcoglycan, Hs01087180_m1 and fly ribosomal protein L32, Dm02151827_g1 for endogenous control).

Techniques:

Journal: PLoS ONE

Article Title: Serial Examination of an Inducible and Reversible Dilated Cardiomyopathy in Individual Adult Drosophila

doi: 10.1371/journal.pone.0007132

Figure Lengend Snippet: (A) Temperature shift from 18°C to 26°C causes the induction of S151A-SGCD expression and subsequent deterioration in cardiac function. At 96 hours post induction, flies expressing S151A-SGCD demonstrate an enlargement in EDD and ESD with a resultant impairment in FS. At 96 hours, a temperature shift back to 18°C results in a repression of S151A-SGCD expression and subsequent improvement in cardiac function with return of EDD, ESD and FS to near baseline. A similar level of wt-SGCD expression after temperature shift from 18°C to 26°C does not result in deterioration in cardiac function. Each graph represents the summary data for serial OCT measurements of EDD, ESD and FS and are expressed as the mean +/− SE (n = 16 for wt-SGCD and n = 16 for S151A-SGCD). * P<0.05 for time point measurements compared to baseline and # P<0.05 for measurements between wt-SGCD and S151A-SGCD at each time point. (B) Representative serial OCT m-modes in individual flies expressing S151A-SGCD or wt-SGCD at the indicated times and temperatures. The arrows indicate EDD when the fly heart is fully relaxed and ESD when the fly heart if fully contracted. A 125 micron standard and 1 second scale bar is shown.

Article Snippet: Applied Biosystems Taqman Gene expression assays were used to perform quantitative (real time) RT-PCR (human delta-sarcoglycan, Hs01087180_m1 and fly ribosomal protein L32, Dm02151827_g1 for endogenous control).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Serial Examination of an Inducible and Reversible Dilated Cardiomyopathy in Individual Adult Drosophila

doi: 10.1371/journal.pone.0007132

Figure Lengend Snippet: (A).Temperature shift from 18°C to 26°C results in deterioration in cardiac function and induction of S151A-SGCD expression. After a temperature shift to 26°C, the induction of transgene expression precedes the development of dilated cardiomyopathy by 48 hours as demonstrated by QRT-PCR measurements of S151A-SGCD mRNA levels compared to cardiac function by serial OCT. A second temperature shift back to 18°C represses S151A-SGCD mRNA expression and results in restoration of cardiac function by 48 hours. We performed three independent experiments using different batches of flies each time. The summary data for QRT-PCR at the indicated times and temperatures are expressed as the mean +/− SE of three independent experiments, each performed in triplicate. * p<0.05 for S151A-SGCD expression at indicted time vs. baseline; # p<0.05 for wt-SGCD expression at the indicated time points vs. baseline; Δ p<0.05 for wt-SGCD vs. S151A-SGCD at the 48 hour time point. (B–D) Summary data for the time course of S151A-SGCD expression (closed circles) vs. cardiac measurements (closed squares) based on serial measurements of EDD, ESD, and FS as described in .

Article Snippet: Applied Biosystems Taqman Gene expression assays were used to perform quantitative (real time) RT-PCR (human delta-sarcoglycan, Hs01087180_m1 and fly ribosomal protein L32, Dm02151827_g1 for endogenous control).

Techniques: Expressing, Quantitative RT-PCR