human kidney epithelial hek293 cells  (ATCC)

Journal: Viruses

Article Title: Unveiling the Role of TMPRSS2 in the Proteolytic Activation of Pandemic and Zoonotic Influenza Viruses and Coronaviruses in Human Airway Cells.

doi: 10.3390/v16111798

Figure Lengend Snippet: Figure 5. TMPRSS2 dependency of H1N1/1918, H12, H13 and H17. (A) Sequence alignment of the cleavage sites from different HAs. (B) HEK293 cells transiently expressing the H1/1918 and human TMPRSS2 or HAT, or murine TMPRSS4 or TMPRSS13 were subjected to SDS-PAGE at 48 h p.t. Western blot analysis was performed with an antibody targeting H1. Tubulin was stained as a loading control. The result shown is representative of three independent experiments. EV = empty vector. (C) Calu-3 cells were treated with 25 µM T-ex5 for 24 h before being infected with rH1N1/1918 at an MOI of 0.001. Viral titers in the supernatant sampled at the indicated time points were determined via TCID50 endpoint titration. The data shown are the mean + SD of three individual experiments. The dotted lines indicate the LOD. (D) The cell lysates and supernatants (SN) of Calu-3 cells treated with 25 µM of T-ex5 for 24 h and then infected with rH1N1/1918 at an MOI of 1 for another 24 h were subjected to SDS-PAGE, and an antibody targeting H1 was used for the Western blot analysis. ß-actin was used as a control. The result is representative of three independent experiments. (E) Calu-3 cells pre-treated with 25 µM were infected with recombinant viruses containing avian H12 or H13 or bat-derived H17 and sharing the other seven gene segments of SC35M at a low MOI of 0.01–0.0001. The supernatant of infected cells was harvested at indicated time points and the viral titers were determined via an immune plaque assay. The experiments were performed in triplicate and the data shown are mean + SD. The dotted lines indicate the LOD. (C,E) Statistically significant differences in viral titers at each time point compared to untreated controls were determined by performing unpaired t-tests. ns = not significant, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.

Article Snippet: HeLa cervical epithelial cells (ATCC, CCL-2), HEK293 human embryonic kidney epithelial cells (ATCC, CRL-1573) and Madin–Darby canine kidney II (MDCKII) cells were grown in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% FBS, 1% glutamine and 1% antibiotics.

Techniques: Sequencing, Expressing, SDS Page, Western Blot, Staining, Control, Plasmid Preparation, Infection, Titration, Recombinant, Derivative Assay, Plaque Assay

Journal: Drug Delivery

Article Title: Biodegradable silica nanoparticles for efficient linear DNA gene delivery

doi: 10.1080/10717544.2024.2385376

Figure Lengend Snippet: p-DNA@SiO 2 particles. (a) TEM images of p-DNA#4 particles with 4.5 µg of plasmid H2B:YFP DNA. (b) Particle diameter (mean ± SD) measured by TEM within a scatter dot plot comparing p-DNA#1-6 ( n = 100). Values for p-DNA#5 and p-DNA#6 are not included due to the presence of aggregates (see Figures S8–S9 ). (*)(**) p-DNA#5-6 not included due to the presence of aggregates (see Figures S5–S6 ). (c) DLS characterization of p-DNA#1-6 bar chart of Z-size (mean ± SD, left Y axis, n = 3) and dot plot in red showing the polydispersity index (PDI) (mean ± SD, right Y axis, n = 3). (d) Flow cytometry quantification of the transfection efficiency (mean fluorescence intensity) of the p-DNA#4 DNA@SiO 2 particles and a representative confocal microscopy image of HEK 293 cells expressing the recombinant H2B:YFP protein 72 h after transfection. Data are shown as the mean ± SD of 3 experimental replicas ( n = 10,000 cells/replica, t- test, * p < 0.05, and *** p < 0.001).

Article Snippet: HEK 293 (Human Kidney Epithelial Cells, RRID:CVCL_0045) cells were acquired in Innoprot (Innovative Technologies in Biological Systems, Derio, Spain).

Techniques: Plasmid Preparation, Flow Cytometry, Transfection, Fluorescence, Confocal Microscopy, Expressing, Recombinant

Journal: Drug Delivery

Article Title: Biodegradable silica nanoparticles for efficient linear DNA gene delivery

doi: 10.1080/10717544.2024.2385376

Figure Lengend Snippet: l-DNA@SiO 2 particles. (a) TEM images of particles l-DNA#4 with 4.5 µg of linear H2B:YFP DNA. (b) Particle diameter (mean ± SD) measured by TEM within a scatter dot plot comparing l-DNA#1-5 ( n = 100). (*) l-DNA#5 is not included due to the presence of aggregates (see Figure S11 ). (c) l-DNA#1-6 bar chart of Z-size (mean ± SD, left Y axis, n = 3) and dot plot in red showing polydispersity index (PDI) (mean ± SD, right Y axis, n = 3). (d) Flow cytometry quantification of the transfection efficiency (mean fluorescence intensity) of the l-DNA#4 DNA@SiO 2 particles and a representative confocal microscopy image of HEK 293 cells expressing the recombinant H2B:YFP protein 72 h after transfection. Data are shown as the mean ± SD of 3 experimental replicas ( n = 10,000 cells/replica, t- test, * p < 0.05, and *** p < 0.001).

Article Snippet: HEK 293 (Human Kidney Epithelial Cells, RRID:CVCL_0045) cells were acquired in Innoprot (Innovative Technologies in Biological Systems, Derio, Spain).

Techniques: Flow Cytometry, Transfection, Fluorescence, Confocal Microscopy, Expressing, Recombinant

Journal: Drug Delivery

Article Title: Biodegradable silica nanoparticles for efficient linear DNA gene delivery

doi: 10.1080/10717544.2024.2385376

Figure Lengend Snippet: Quantification of H2B:YFP protein expression (mean fluorescence intensity) using Lipofectamine TM 2000 and DNA@SiO 2 system with p-DNA and l-DNA in HEK 293 cells. Data are shown as the mean ± SD of 3 experimental replicas ( n = 10,000 cells/replica, t- test, * p < 0.05, and *** p < 0.001).

Article Snippet: HEK 293 (Human Kidney Epithelial Cells, RRID:CVCL_0045) cells were acquired in Innoprot (Innovative Technologies in Biological Systems, Derio, Spain).

Techniques: Expressing, Fluorescence