human embryonic kidney 293 t hek293t cell line  (Procell Inc)

Journal: Journal of Translational Medicine

Article Title: Nicotine restores olfactory function by activation of prok2R/Akt/FoxO3a axis in Parkinson’s disease

doi: 10.1186/s12967-024-05171-1

Figure Lengend Snippet: Nicotine treatment mitigates MPP + -induced apoptosis in HEK293T cells. a Safety evaluation of nicotine using the Cell Counting Kit-8 (CCK 8) assay. n = 6 in each group. b Damage assessment of MPP + using the CCK8 assay. n = 6 in each group. c Improvement in cell viability owing to nicotine treatment is concentration dependent. n = 6 in each group. d The bright field and live/dead staining images of different treatment groups. Scale bar = 100 μm. e , f Apoptotic flow cytometry and quantitative analyses of the apoptotic cell percentage. n = 3 in each group. g Transmission electron microscopy images of HEK293T cells. The phagosome (red triangle labelled) in the nicotine only treated group indicates engulfment of small molecules, and the HEK293T cells of the nicotine + MPP + group are intact. MPP + treated HEK293T cells manifest early apoptosis accompanied by nuclear fragmentation, condensation, and membrane integrity. These cells show ruptured membranes and the disappearance of normal cellular structures during late apoptosis. Scale bar = 5 μm. Significant differences are shown by *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: Human embryonic kidney 293 T (HEK293T) cell line and primary mouse olfactory neurons (CP-M145) were purchased from Procell (Wuhan, China).

Techniques: Cell Counting, CCK-8 Assay, Concentration Assay, Staining, Flow Cytometry, Transmission Assay, Electron Microscopy, Membrane

Journal: Journal of Translational Medicine

Article Title: Nicotine restores olfactory function by activation of prok2R/Akt/FoxO3a axis in Parkinson’s disease

doi: 10.1186/s12967-024-05171-1

Figure Lengend Snippet: Nicotine activates the prok2R/Akt/FoxO3a axis and exhibits anti-apoptotic effects against MPP + in the HEK293T cell line. Western blot (WB) images ( a ) and semi-quantitative analysis ( b – d ) of prok2R/Akt/FoxO3a axis. Confocal laser scanning microscopy (CLSM) images of fluorescein isothiocyanate (FITC)-labeled prok2R ( e ) and semi-quantitative analysis of fluorescence intensity ( f ). Scale bar = 10 μm. The expression levels of anti-apoptotic protein (Bcl-2), pro-apoptotic protein (Bax), and apoptotic executor protein (Cl-casp3) detected using WB ( g – j ). k , l CLSM of FITC-labeled Cl-casp3 indicating decreased expression levels after nicotine treatment against MPP + damage. Scale bar = 10 μm. Significant differences are shown by *p < 0.05, **p < 0.01, ***p < 0.001. n = 3 in each group

Article Snippet: Human embryonic kidney 293 T (HEK293T) cell line and primary mouse olfactory neurons (CP-M145) were purchased from Procell (Wuhan, China).

Techniques: Western Blot, Confocal Laser Scanning Microscopy, Labeling, Fluorescence, Expressing

Journal: Journal of Translational Medicine

Article Title: Nicotine restores olfactory function by activation of prok2R/Akt/FoxO3a axis in Parkinson’s disease

doi: 10.1186/s12967-024-05171-1

Figure Lengend Snippet: Prok2R expression levels modulate activation of the Akt/FoxO3a axis. Western blot (WB) images of prok2R/Akt/FoxO3a in prok2R + HEK293T cell lines ( a – d ). WB images of prok2R/Akt/FoxO3a in prok2R − HEK293T cell lines ( e – h ). The prok2R/Akt/FoxO3a axis expression levels in the prok2R vector , prok2R + , prok2R + + DMSO, and prok2R + + ipatasertib groups ( i – l ). Apoptosis-related proteins expressed in different treatment groups ( m – p ). Significant differences are shown by *p < 0.05, **p < 0.01, ***p < 0.001. n = 3 in each group

Article Snippet: Human embryonic kidney 293 T (HEK293T) cell line and primary mouse olfactory neurons (CP-M145) were purchased from Procell (Wuhan, China).

Techniques: Expressing, Activation Assay, Western Blot, Plasmid Preparation

Journal: Journal of Translational Medicine

Article Title: Nicotine restores olfactory function by activation of prok2R/Akt/FoxO3a axis in Parkinson’s disease

doi: 10.1186/s12967-024-05171-1

Figure Lengend Snippet: Prok2R + HEK293T cells alleviate MPP + -induced apoptosis through the preservation of prok2R/Akt/FoxO3a axis activity. The activity of prok2R/Akt/FoxO3a in different treatment groups (prok2R vector , prok2R vector + MPP + , prok2R + , prok2R + + MPP + ) revealed using the western blot (WB) assay ( a – d ). Apoptosis-related proteins indicated using the WB assay ( e – h ). Confocal laser scanning microscopy images of Cy3-labeled Cl-casp3 in each group ( i , j ). Scale bar = 10 μm. Significant differences are shown by *p < 0.05, **p < 0.01, ***p < 0.001. n = 3 in each group

Article Snippet: Human embryonic kidney 293 T (HEK293T) cell line and primary mouse olfactory neurons (CP-M145) were purchased from Procell (Wuhan, China).

Techniques: Preserving, Activity Assay, Plasmid Preparation, Western Blot, Confocal Laser Scanning Microscopy, Labeling

Journal: Signal Transduction and Targeted Therapy

Article Title: Exosomes secreted from cardiomyocytes suppress the sensitivity of tumor ferroptosis in ischemic heart failure

doi: 10.1038/s41392-023-01336-4

Figure Lengend Snippet: Direct interaction between miR-22-3p and ferroptosis-related gene ACSL4. a The sequence of miR-22-3p and the potential binding site of ACSL4 mRNA; b ACSL4 gene expression in lung cancer tissues and paracancerous tissues from TCGA database; c , d The protein expression levels of ACSL4 were determined by western blot in ( c ) subcutaneous xenograft tissues was determined ( N = 3/group) and in ( d ) LLC cells after transfecting with miR-22-3p and NC in the presence of erastin ( N = 5 independent experiments); e Luciferase activity assay was performed to confirm the ACSL4 mRNA was directly bound to miR-22-3p in HEK-293T cells ( N = 3 independent experiments); f LLC cells were co-transfected with ACSL4 overexpression/empty constructs and miR-22-3p mimics/NC for 24 h. Representative images and quantitative results of LLC cancer cell colonies ( N = 3 independent experiments); g Complementarity between miR-22-3p seed sequence and the ACSL4 mRNA, the miRNA-masking antisense (ODN-22-3p) was designed to be fully complementary to the miR-22-3p targeting sequence on ACSL4; h – j LLC cells were transfected with miR-22-3p mimics and ODN in the presence of erastin for 24 h. h Analysis of lipid-ROS using C11 BODIPY 581/591 fluorescence staining (Bar: 40 μm), Red, non-oxidized form of C11-BODIPY; Green, oxidized form of C11-BODIPY. Each data point represents the ratio of oxidized C11 to non-oxidized C11 signal ( N = 10 from 3 independent experiments); i The protein expression levels of ACSL4 were determined by western blot ( N = 3 independent experiments); j Representative images and quantitative results of LLC cancer cell colonies ( N = 3 independent experiments). Data are expressed as mean ± SEM; * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Mouse Lewis lung carcinoma cell line (LLC), osteoblast cell line from the bone of a mouse with osteosarcoma (K7M2), and human embryonic kidney 293 T (HEK-293T) cell line were cultured in DMEM supplemented with 10% FBS (Biological Industries, Israel).

Techniques: Sequencing, Binding Assay, Expressing, Western Blot, Luciferase, Activity Assay, Transfection, Over Expression, Construct, Fluorescence, Staining