Journal: International Journal of Molecular Sciences

Article Title: Unravelling Novel SCN5A Mutations Linked to Brugada Syndrome: Functional, Structural, and Genetic Insights

doi: 10.3390/ijms242015089

Figure Lengend Snippet: Electrophysiological characterization of Nav1.5 constructs. Representative current traces and their biophysical properties measured via the patch clamp approach in HEK293 cells overexpressing the constructs as indicated: WT in black ( A ), WT/p.A344S or p.A344S in blue ( B ), WT/p.N347K or p.N347K in green ( C ), and WT/p. D349N or p.D349N in red ( D ). Panels B, C, and D present from left to right: families of current traces, current–voltage relationship showing the reduction in the mean current density recorded in the presence of the mutations in heterozygous or homozygous conditions with respect to the WT channel, fitting the data with the Boltzmann equation (see ) provided midpoint activation and inactivation potential reported in , recovery from inactivation (see for the kinetic parameters), and the kinetics of the development of the intermediate inactivation. Data are shown as mean ± SEM.

Article Snippet: The HEK293 (human embryonic kidney 293) cell line was obtained from ATCC.

Techniques: Construct, Patch Clamp, Activation Assay

Journal: International Journal of Molecular Sciences

Article Title: Unravelling Novel SCN5A Mutations Linked to Brugada Syndrome: Functional, Structural, and Genetic Insights

doi: 10.3390/ijms242015089

Figure Lengend Snippet: Localization of Nav1.5 WT and mutants transiently transfected in HEK293 cells. Representative immunofluorescence confocal images showing the plasma membrane and cytosolic localization of WT Nav1.5 protein ( A ), along with its p.A344S ( B ), p.N347K ( C ), and p.D349D ( D ) mutants. Nav1.5 is labeled in green. Scale bar: 10 µm.

Article Snippet: The HEK293 (human embryonic kidney 293) cell line was obtained from ATCC.

Techniques: Transfection, Immunofluorescence, Membrane, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Unravelling Novel SCN5A Mutations Linked to Brugada Syndrome: Functional, Structural, and Genetic Insights

doi: 10.3390/ijms242015089

Figure Lengend Snippet: Effect of mexiletine on the mutated constructs. ( A ) Representative current traces of mutated channels recorded after mexiletine incubation in response to a depolarizing stimulus at −20 mV in HEK293 cells expressing the constructs as indicated: WT in black (for reference), p.A344S in blue, p.N347K in green, and p.D349N in red. ( B ). Scattered plot showing the distribution of the current density values obtained in absence (filled symbols) and in presence (empty symbols) of mexiletine. The black line represents the average values (see ). ( C ) Fitting the data with the Boltzmann equation (see the ) provided midpoint activation potential, as reported in . Data are shown as mean ± SEM.

Article Snippet: The HEK293 (human embryonic kidney 293) cell line was obtained from ATCC.

Techniques: Construct, Incubation, Expressing, Activation Assay

Journal: Cellular and Molecular Life Sciences

Article Title: Intracellular FGF1 protects cells from apoptosis through direct interaction with p53

doi: 10.1007/s00018-023-04964-9

Figure Lengend Snippet: Transiently expressed FGF1 protects cell against apoptosis only in p53-positive cells. a , b MCF7, BJ and HEK 293 cell lines expressing WT p53 (a) and p53-negative cell lines G292 and PC3 b were transiently transfected with myc-FGF1_pcDNA3.1 or control pcDNA3.1 vectors. 48 h after transfection, cells were treated with treated with 1 μM staurosporine, 10 μM anisomycin or 5 μM actinomycin D in the presence of FGFR kinase inhibitor (100 nM PD173074) for 24 h. Cell viability was measured using Presto Blue reagent. Graphs show means ± SD from three independent experiments. Statistical significance: *p < 0.05, **p < 0.01

Article Snippet: The primary human foreskin fibroblast BJ cell line (CRL-2522), human osteosarcoma U2OS cell line (HTB-96), human breast cancer MCF7 cell line (HTB-22), human osteosarcoma G292 cell line, human prostate cancer PC3 cell line (CRL-1435) and human embryonic kidney HEK-293 cell line (CRL-1573) were from ATCC (VA, USA).

Techniques: Expressing, Transfection