human embryonic kidney cell line hek293t  (ATCC)

Journal: bioRxiv

Article Title: Enhancing the biocorrosion resistance and biocompatibility of Aluminium substrates using Graphene Oxide-PEDOT:PSS Hybrid Coating

doi: 10.1101/2025.09.29.679258

Figure Lengend Snippet: Comparison of cell viability, assessed by MTT method in the DMEM culture medium, after exposure to ( A ) Al_GO/P sample extracts by indirect contact assay and ( B ) after direct contact with the Al_GO/P samples; at ( i ) 24-hour and ( ii ) 48-hour. Data are expressed as % values of result obtained in comparison to control cells (HEK293T) not exposed to materials or material extracts. Results from indirect contact assay showed Al_GO/P substrates maintained optimal biocompatibility compared to bare Al over 48 hours. While direct contact assay indicated high cell viability for Al_GO50P and Al_GO100P, with Al_GO250P showing minor viability changes at 48 hours, while significant decreases were observed for Al_GO500P and Al_GO1000P over 48 hours. Error bars represent ±1 standard deviation (SD) from mean forthree independent biological replicates (n=3). Statistical significance was assessed using one-way ANOVA; specific p-values of <0.05, <0.01, <0.001, and <0.0001 were indicated as *,**,***, and ****, respectively.

Article Snippet: The Human Embryonic Kidney (HEK293T) cell line [ATCC CRL-11268] was used.

Techniques: Comparison, Control, Standard Deviation

Journal: Virologica Sinica

Article Title: Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway

doi: 10.1016/j.virs.2025.08.004

Figure Lengend Snippet: UXT-V2 inhibits HSV-2 replication via NF-κB independent pathway. A ARPE-19 cells were transfected with an empty vector or different doses of plasmids expressing His-UXT-V1 (0.5 or 1 μg) or His-UXT-V2 (0.5 or 1 μg), followed by infection with HSV-2 at an MOI of 0.2 PFU/cell at 6 hpt for 18 h. The virus yields were measured by plaque assay. B HeLa cells were transfected with 0.3 μg PRD(III-I) 4 -Luc or NF-κB–Luc reporter plasmid along with 1.5 μg empty vector or 1.5 μg plasmid expressing His-UXT-V1 or His-UXT-V2, followed by infection with HSV-2 at an MOI of 0.2 PFU/cell at 6 hpt for 18 h. The luciferase activity was measured to determine the activation fold. HEK293T cells were transfected with 0.3 μg NF-κB-Luc reporter plasmid along with an empty vector or 1 μg plasmid expressing His-UXT-V2 for 24 h, followed by treatment with or without 10 ng/mL TNF-α for 6 h. Luciferase activity was measured to determine the activation fold. C HEK293T cells were transfected with 0.3 μg NF-κB-Luc reporter plasmid along with an empty vector or 1 μg plasmid expressing His-UXT-V2 for 24 h, followed by treatment with or without 10 ng/mL TNF-α for 6 h. Luciferase activity was measured to determine the activation fold. D BHK-21 cells were transfected with an empty vector or different doses of plasmids expressing His-UXT-V1 (0.5 or 2 μg) or His-UXT-V2 (0.5 or 2 μg), followed by infection with JEV at an MOI of 0.5 PFU/cell at 6 hpt. The supernatants and cells were harvested, and the virus yields were measured by plaque assay at 24 hpi. E BHK-21 cells were transfected with an empty vector or 1.5 μg plasmid expressing His-UXT-V2, followed by infection with JEV at an MOI of 0.5 PFU/cell at 6 hpt. The supernatants and cells were harvested, and the virus yields were measured by plaque assay at 8, 16, 24 and 36 hpi. F Vero E6 cells were transfected with an empty vector or different doses of plasmids expressing His-UXT-V1 (0.5 or 2 μg) or His-UXT-V2 (0.5 or 2 μg), followed by infection with ZIKV at an MOI of 0.5 PFU/cell at 6 hpt. The supernatants and cells were harvested and the virus yields were measured by plaque assay at 24 hpi. G Vero E6 cells were transfected with an empty vector or 1.5 μg plasmid expressing His-UXT-V2, followed by infection with ZIKV at an MOI of 0.5 PFU/cell at 6 hpt. The supernatants and cells were harvested, and the virus yields were measured by plaque assay at 8, 16, 24 and 36 hpi. H Vero E6 cells transfected with the control or the two UXT shRNAs, respectively, were infected with HSV-2 at an MOI of 0.2 PFU/cell for 24 h. The virus yields were measured by plaque assay. For graphs, data shown are mean ± SD of three independent experiments with each condition performed in triplicate (∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; ns, not significant). For images, one representative experiment out of three is shown.

Article Snippet: Human embryonic kidney 293T (HEK293T) cell line, baby hamster kidney cell line BHK-21, African green monkey kidney cell line Vero and Vero E6, and human cervical epithelial cell line HeLa were purchased from the American Type Culture Collection.

Techniques: Transfection, Plasmid Preparation, Expressing, Infection, Virus, Plaque Assay, Luciferase, Activity Assay, Activation Assay, Control

Journal: Virologica Sinica

Article Title: Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway

doi: 10.1016/j.virs.2025.08.004

Figure Lengend Snippet: UXT-V2 downregulates the expression of HSV-2 gB. A HSV-2 proteins fished out by LC-MS/MS analysis. Figure was created using ggplot2. B HEK293T cells were co-transfected with 2 μg empty vector or 2 μg His-UXT-V2 together with 2 μg plasmid expressing Flag-gB. At 24 hpi, cells were harvested and lysed, and the extracts were subjected to IP using the anti-Flag MAb or control IgG. C HeLa cells were infected with or without HSV-2 at an MOI of 0.2 PFU/cell. At 24 hpi, cells were harvested and lysed, and the extracts were subjected to IP using the anti-gB MAb. D HEK293T cells were co-transfected with 1.5 μg empty vector or 1.5 μg plasmid expressing His-UXT-V2 together with 0.5 μg plasmid expressing various Flag-tagged HSV-2 proteins for 24 h. The expression of Flag-tagged HSV-2 proteins were measured by Western blotting. E HEK293T cells were co-transfected with 1.5 μg empty vector or 1.5 μg plasmid expressing His-UXT-V2 together with 0.5 μg plasmid expressing Flag-gB for 12 or 18 h. Cells were harvested and the expression of Flag-gB was detected by Western blotting. F HEK293T cells were co-transfected with 1.5 μg empty vector or 1.5 μg plasmid expressing His-UXT-V1 or His-UXT-V2 together with 0.5 μg plasmid expressing Flag-gB for 12 or 18 h. Cells were harvested and the expression of Flag-gB was measured by Western blotting. G ARPE-19 cells were transfected with 60 nM NC or #4 (UXT-V2 siRNA), followed by transfection with an empty vector or different doses of plasmids expressing Flag-gB (0.5 or 2 μg) for 24 h. Cells were harvested and the expression of Flag-gB was detected by Western blotting. H HeLa cells were transfected with an empty vector or 2 μg plasmid expressing His-UXT-V2, followed by infection with HSV-2 at an MOI of 5 PFU/cell. At 6 or 8 hpi, cells were harvested, and the expression level of HSV-2 gB or gD was analyzed by Western blotting. I WT and UXT-V2-KO HeLa cells were infected with HSV-2 at an MOI of 5 PFU/cell. At 6 or 8 hpi, cells were harvested and the expression level of HSV-2 gB or gD was measured by Western blotting. J HeLa cells were co-transfected with an empty vector or different doses of plasmids expressing His-UXT-V2 (0.5 or 2 μg) together with 0.5 μg plasmid expressing Flag-gB, followed by infection with HSV-2 at an MOI of 0.2 PFU/cell at 6 hpt for 18 h. Virus yields were measured by plaque assay. The graph shows compensating efficiency of gB on HSV-2 yields. For graphs, data shown are mean ± SD of three independent experiments with each condition performed in triplicate (∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001; ns, not significant). For images, one representative experiment out of three is shown.

Article Snippet: Human embryonic kidney 293T (HEK293T) cell line, baby hamster kidney cell line BHK-21, African green monkey kidney cell line Vero and Vero E6, and human cervical epithelial cell line HeLa were purchased from the American Type Culture Collection.

Techniques: Expressing, Liquid Chromatography with Mass Spectroscopy, Transfection, Plasmid Preparation, Control, Infection, Western Blot, Virus, Plaque Assay

Journal: Virologica Sinica

Article Title: Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway

doi: 10.1016/j.virs.2025.08.004

Figure Lengend Snippet: UXT-V2 induces gB degradation through proteasomal pathways. A HEK293T cells were co-transfected with 1.5 μg empty vector or 1.5 μg plasmid expressing His-UXT-V2 together with 0.5 μg plasmid expressing Flag-gB for 12 h or 18 h. Cells were harvested and the transcripts of gB were detected by relative quantitative PCR. B HEK293T cells were co-transfected with 1.5 μg empty vector or 1.5 μg plasmid expressing His-UXT-V2 together with 0.5 μg plasmid expressing Flag-gB in the absence or presence of CQ (20 or 100 μM) or MG132 (1 or 2 μM). At 24 hpt, the expression of Flag-gB was detected by Western blotting. C HEK293T cells were transfected with 1 μg plasmid expressing Flag-gB in the absence or presence of CQ (20 or 100 μM), 3-MA (5 or 20 μM) or MG132 (1 or 2 μM). At 24 hpt, the expression of Flag-gB was detected by Western blotting. D HEK293T cells were co-transfected with 1.5 μg empty vector or 1.5 μg plasmid expressing His-UXT-V2 together with 0.5 μg plasmid expressing Flag-gB in the absence or presence of CQ (20 or 100 μM), 3-MA (5 or 20 μM) or MG132 (1 or 2 μM). At 24 hpt, the expression of Flag-gB was detected by Western blotting. For graphs, data shown are mean ± SD of three independent experiments with each condition performed in triplicate (ns, not significant). For images, one representative experiment out of three is shown.

Article Snippet: Human embryonic kidney 293T (HEK293T) cell line, baby hamster kidney cell line BHK-21, African green monkey kidney cell line Vero and Vero E6, and human cervical epithelial cell line HeLa were purchased from the American Type Culture Collection.

Techniques: Transfection, Plasmid Preparation, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Virologica Sinica

Article Title: Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway

doi: 10.1016/j.virs.2025.08.004

Figure Lengend Snippet: UXT-V2 promotes K48-linked polyubiquitination of HSV-2 gB. A HEK293T cells were co-transfected with 0.5 μg empty vector or different doses of plasmids expressing His-UXT-V2 (0.2, 0.5 or 1.5 μg) together with 0.5 μg plasmid expressing HA-Ub. At 24 hpt, the expression of His-UXT-V2 and ubiquitinated cellular proteins was detected by Western blotting. B HEK293T cells were transfected with 0.5 μg empty vector or different doses of plasmids expressing His-UXT-V2 (0.2, 0.5 or 1.5 μg). At 24 hpt, the expression of His-UXT-V2 and ubiquitinated cellular proteins was detected by Western blotting. C HEK293T cells were co-transfected with 5 μg empty vector or 5 μg plasmid expressing His-UXT-V2 together with 4 μg plasmid expressing Flag-gB for 24 h. Cells were immunoprecipitated with the anti-Flag MAb. D HEK293T cells were co-transfected with 2 μg empty vector or 2 μg plasmid expressing His-UXT-V2 together with 2 μg plasmid expressing Flag-gB and 2 μg plasmid expressing HA-Ub, HA-K48 or HA-K63 for 24 h. Cells were immunoprecipitated with the indicated antibodies. E HeLa cells were co-transfected with 2 μg empty vector or 2 μg plasmid expressing His-UXT-V2 together with 2 μg plasmid expressing HA-Ub, HA-K48 or HA-K63, followed by infection with HSV-2 at an MOI of 2 PFU/cell at 6 hpt for 18 h. Cells were immunoprecipitated with the indicated antibodies. For images, one representative experiment out of three is shown.

Article Snippet: Human embryonic kidney 293T (HEK293T) cell line, baby hamster kidney cell line BHK-21, African green monkey kidney cell line Vero and Vero E6, and human cervical epithelial cell line HeLa were purchased from the American Type Culture Collection.

Techniques: Transfection, Plasmid Preparation, Expressing, Western Blot, Immunoprecipitation, Infection

Journal: Virologica Sinica

Article Title: Ubiquitously expressed transcript isoform 2 (UXT-V2) restricts HSV-2 replication by targeting glycoprotein B for degradation through ubiquitin-proteasome pathway

doi: 10.1016/j.virs.2025.08.004

Figure Lengend Snippet: UXT-V2 facilitates TRIM21-dependent proteasomal degradation of gB. A E3 ubiquitin ligases fished out by LC-MS/MS analysis. Figure was created using ggplot2. B HEK293T cells were co-transfected either with 2 μg empty vector or 2 μg HA-TRIM21 expression plasmid, along with 2 μg plasmid expressing Flag-gB. At 24 hpt, cells were harvested and lysed, followed by IP using the anti-Flag MAb or control IgG. C The expression of TRIM21 in sgControl and sgTRIM21 HeLa cells was assessed by immunoblot analysis. D HeLa or TRIM21-KO HeLa cells were co-transfected with either 2 μg empty vector or 2 μg plasmid expressing His-UXT-V2, along with 2 μg plasmid expressing Flag-gB and 2 μg plasmid expressing HA-K48. At 24 hpt, cells were harvested and lysed, followed by IP using the anti-Flag Mab or control IgG. For images, one representative experiment out of three is shown.

Article Snippet: Human embryonic kidney 293T (HEK293T) cell line, baby hamster kidney cell line BHK-21, African green monkey kidney cell line Vero and Vero E6, and human cervical epithelial cell line HeLa were purchased from the American Type Culture Collection.

Techniques: Ubiquitin Proteomics, Liquid Chromatography with Mass Spectroscopy, Transfection, Plasmid Preparation, Expressing, Control, Western Blot