hek 293 cells  (AMS Biotechnology)

Journal: Translational Psychiatry

Article Title: Immunoglobulin G modulation of the melanocortin 4 receptor signaling in obesity and eating disorders

doi: 10.1038/s41398-019-0422-9

Figure Lengend Snippet: a Representative images of hMC4R GFP+ HEK 293 cells (green) 30 min after application of DyLight 550®-labeled (red) α-MSH affinity-purified IgG from eating disorder (anorexia nervosa, n = 9; bulimia nervosa, n = 7; binge eating disorder, n = 7), obese ( n = 10), and Ctrl ( n = 9) preincubated or not with α-MSH. Quantification of DyLight 550®-positive spots in hMC4R+ HEK 293 cells ( n = 50/group): b at the membrane; c intracellularly (cytosolic), and d ratios of cytosolic/membrane staining. Affinity kinetics properties of α-MSH/IgG IC for hMC4R + HEK 293 cells including e dissociation equilibrium constant (KD); f association rate (ka), and g dissociation rate (kd). Data are means ± s.e.m. Kruskal–Wallis test with Dunns’ post-tests ( b – d , f , g ) or analysis of variance with Tukey’s post-test ( e ), *** p < 0.001, ** p < 0.01, * p < 0.05

Article Snippet: MC4R-expressing HEK 293 cells (AMS Biotechnology, Abingdon, UK) were cultured in glass bottom Petri dishes (MatTek, ≈250,000 cells/dish).

Techniques: Labeling, Affinity Purification, Staining

Journal: Translational Psychiatry

Article Title: Immunoglobulin G modulation of the melanocortin 4 receptor signaling in obesity and eating disorders

doi: 10.1038/s41398-019-0422-9

Figure Lengend Snippet: a cAMP dose–response curves to α-MSH alone or α-MSH/IgG IC formed by IgG pooled in patents and control groups and adjusted to α-MSH-reactive IgG plasma levels of controls. cAMP dose–response curves to α-MSH preincubated or not with individual total IgG and corresponding EC50 ( b ) and maximal cAMP production ( c ). d Control experiments including cAMP dose–response curves to α-MSH by MC4R-expressing and non-expressing control HEK 293 cells and to α-MSH 1–4 peptide by MC4R-expressing cells ( n = 4). e cAMP dose-response curves to α-MSH and IgG from patients and controls without their overnight pre-incubation. f , cAMP dose–response curves to α-MSH alone or α-MSH/IgG IC co-administered (solid line) with agouti-related protein (AgRP; 100 nM) or added after AgRP preincubation (dotted line— n = 2/group) as well as in g cAMP maximal response. h , i cAMP dose–response curves of α-MSH preincubated with h purified total IgG from patients and controls depleted for α-MSH-reactive IgG ( n = 3/group) and i affinity-purified α-MSH-reactive IgG ( n = 6/group); j EC 50 and k maximal cAMP production at the plateau. Data are means ± s.e.m. Analysis of variance with Tukey’s post-test ( b , c , g , j ) or Kruskal–Wallis test with Dunns’ post-tests ( m ), *** p < 0.001, ** p < 0.01, * p < 0.05; Mann–Whitney test, $ p < 0.05. a α-MSH ( n = 9), Ctrl, anorexia nervosa (AN), bulimia nervosa (BN), and binge eating disorder (BED; n = 6), obese (OB; n = 4); d HEK 293-hMC4R+ ( n = 5), HEK 293-CTRL ( n = 6); e α-MSH ( n = 3), Ctrl, BN, and BED ( n = 2), AN and OB ( n = 3)

Article Snippet: MC4R-expressing HEK 293 cells (AMS Biotechnology, Abingdon, UK) were cultured in glass bottom Petri dishes (MatTek, ≈250,000 cells/dish).

Techniques: Expressing, Incubation, Purification, Affinity Purification, MANN-WHITNEY

Journal: Stem Cell Research & Therapy

Article Title: Lin28A induces energetic switching to glycolytic metabolism in human embryonic kidney cells

doi: 10.1186/s13287-016-0323-2

Figure Lengend Snippet: Characterisation of Lin28A over-expression in HEK 293 cells. Real-time PCR revealed that Lin28 mRNA expression levels were increased in Lin28A over-expressing cells, n = 3 ( a ). Conversely, Let 7 a, c and g miRNA expression levels were decreased in Lin28 over-expressing cells, n = 3 ( b ). Western blot analysis ( c ) confirmed that Lin28 protein levels were also increased in over-expressing HEK cells ( d ), n = 4. GAPDH and U6 were used as housekeeping genes for mRNA and miRNA expression, respectively. Student’s unpaired t-tests were used to compare between experimental groups. Data are represented as mean ± SEM. HEK human embryonic kidney, miRNA microRNA

Article Snippet: Lin28A over-expressing human embryonic kidney (HEK) 293 cells were obtained from Amsbio, UK.

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Expressing, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: Lin28A induces energetic switching to glycolytic metabolism in human embryonic kidney cells

doi: 10.1186/s13287-016-0323-2

Figure Lengend Snippet: Lin28A increases cellular proliferation and glucose transport in HEK 293 cells. Proliferation studies revealed that cell density was significantly increased at 72 and 96 hour time points in Lin28A over-expressing cells compared to non-transfected (blank) HEK cells, n = 3 ( a ). Radio-labelled glucose transport assay revealed that 2-deoxyglucose transport was also significantly increased in the Lin28A over-expressing cells (P < 0.01, n = 6) ( b ). Student’s unpaired t-tests were used to compare between experimental groups. Data are represented as mean ± SEM. HEK human embryonic kidney

Article Snippet: Lin28A over-expressing human embryonic kidney (HEK) 293 cells were obtained from Amsbio, UK.

Techniques: Expressing, Transfection, Transport Assay