human embryonic kidney 293 t hek293t cells  (Thermo Fisher)

Journal: Communications Biology

Article Title: SunTag-PE: a modular prime editing system enables versatile and efficient genome editing

doi: 10.1038/s42003-025-07893-4

Figure Lengend Snippet: Comparison of SunTag-ePE3 with canonical fused ePE3, Split-ePE3, and MS2-ePE3 at four endogenous loci, including HEK3 locus ( a ), EMX1 locus ( b ), FANCF locus ( c ), and RNF2 locus ( d ) in HEK293T cells. Each locus contains three types of prime editing: nucleotide transversion, insertion, and deletion. Data and error bars indicate the mean and s.d. of three independent biological replicates. Negative controls represent four kinds of PE3 components without pegRNAs. The numbers above the bars represent fold change between different treatment groups.

Article Snippet: Human Embryonic Kidney 293 T (HEK293T) cell line and HeLa cervical cancer cell line were purchased from ATCC.

Techniques: Comparison

Journal: Cell Death & Disease

Article Title: Zipper-interacting protein kinase mediates neuronal cell death and cognitive dysfunction in traumatic brain injury via regulating DEDD

doi: 10.1038/s41419-025-07474-7

Figure Lengend Snippet: A – C Immunoblot analysis of DEDD expression in primary neurons from WT and Zipk +/− mice ( A ), SH-SY5Y cells with ZIPK knockdown ( B ) or overexpression ( C ). D – F DEDD mRNA levels were analyzed by qRT-PCR in WT and Zipk +/− primary neurons ( D ) and SH-SY5Y cells with ZIPK knockdown ( E ) or overexpression ( F ). G HEK293T cells were transiently transfected with HA-vector or HA-ZIPK and Flag-DEDD constructs and subjected to a CHX assay. Representative immunoblot showing DEDD levels at different time points after CHX addition was shown. H SH-SY5Y cells were transiently transfected with HA-vector or HA-ZIPK constructs and then treated with CHX for the indicated duration, followed by immunoblot quantification of DEDD protein levels. NC, negative control. * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant. Unpaired two-tailed Student’s t test was used for analysis.

Article Snippet: Human embryonic kidney 293 T (HEK293T) cells were obtained from the Cell Bank/Stem Cell Core Facility (Shanghai, China), and was maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, New York, NY, USA) supplemented with 10% fetal bovine serum (FBS, PAN-biotech, Aidenbach, Germany).

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Quantitative RT-PCR, Transfection, Plasmid Preparation, Construct, Negative Control, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Zipper-interacting protein kinase mediates neuronal cell death and cognitive dysfunction in traumatic brain injury via regulating DEDD

doi: 10.1038/s41419-025-07474-7

Figure Lengend Snippet: A , B Endogenous binding of ZIPK and DEDD in mouse brain tissues in the Co-IP assay using anti-ZIPK ( A ) or -DEDD ( B ) antibody, respectively. C Representative autoradiography data from the in vitro kinase assay using 32 P-ATP and purified recombinant proteins. GST-MLC and GST were positive and negative controls, respectively. D 32 P-autoradiography data for WT or alanine mutants of DEDD in in vitro kinase assay. E Phosphorylation of DEDD-WT or S9A mutant detected by an anti-pan-phospho-antibody. F Immunoblot analysis of DEDD-WT and S9A mutant levels in HEK293T cells transiently transfected with Flag-DEDD-WT or Flag-DEDD-S9A and HA-ZIPK constructs in the CHX assay. G , H TUNEL assay ( G ) and immunoblot analysis of cle-caspase-3 ( H ) in SH-SY5Y cells expressing Flag-DEDD-WT or Flag-DEDD-S9A and treated with H 2 O 2 . The scale bar is 100 μm. I Expression levels of cle-caspase-3 in SH-SY5Y cells after co-transfection of HA-ZIPK with Flag-DEDD-WT or Flag-DEDD-S9A and treated with H 2 O 2 . * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Unpaired two-tailed Student’s t test was used in ( F ). One-way ANOVA was used in ( G – I ).

Article Snippet: Human embryonic kidney 293 T (HEK293T) cells were obtained from the Cell Bank/Stem Cell Core Facility (Shanghai, China), and was maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, New York, NY, USA) supplemented with 10% fetal bovine serum (FBS, PAN-biotech, Aidenbach, Germany).

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Autoradiography, In Vitro, Kinase Assay, Purification, Recombinant, Mutagenesis, Western Blot, Transfection, Construct, TUNEL Assay, Expressing, Cotransfection, Two Tailed Test

Journal: Frontiers in Veterinary Science

Article Title: A functional SNP of the core promoter region within goat C DC25A gene affects litter size

doi: 10.3389/fvets.2024.1471123

Figure Lengend Snippet: Mechanism diagram. (A) The structure patterns of goat CDC25A gene. (B) The core promoter of goat CDC25A gene was located at position-663 to-237. The electrophoregram on the left displayed that the five fragments were amplified successfully. The bar plots in the middle and on the right displayed the relative luciferase activity of different fragments within CDC25A gene promoter detected in HEK293T and CHO-K1 cells, respectively. (C) One novel SNP (NC_030829.1:g.51731829A > C) within the core promoter of goat CDC25A gene was found, which might change the binding of transcription factors. (D) The novel SNP significantly affected the transcriptional activity of CDC25A gene. (E) Left: associations between the novel SNP and litter size in goat CDC25A gene and the proportion of different types of litter size in individuals with AA, AC and CC genotypes. Right: the proportion of AA, AC and CC genotypes in single-lamb and multi-lamb groups. SBWC goat, Shaanbei white cashmere goat; UTR, untranslated region; CDS, coding sequence; ** p < 0.01. Bars with different letters (a, b, c, d) means p < 0.05.

Article Snippet: Chinese hamster ovary K1 (CHO-K1) cells and Human embryonic kidney 293 T (HEK293T) cells were cultured in complete medium consisting of Ham’s F 12 nutrient medium (Sangon Biotech, Xi’an, China), 10% fetal bovine serum (Zeta life, America) and 1% penicillin–streptomycin (Gibco, America) at 37°C in a humidified 5% CO 2 atmosphere ( – ).

Techniques: Amplification, Luciferase, Activity Assay, Binding Assay, Sequencing