phiv luc  (Addgene inc)

Journal: bioRxiv

Article Title: The proton-activated chloride channel inhibits SARS-CoV-2 spike protein-mediated viral entry through the endosomal pathway

doi: 10.1101/2025.03.12.642872

Figure Lengend Snippet: A) HEK 293T ACE2 cells (CTRL) or PAC inducible cells (PAC) treated with SARS-CoV-2, SARS CoV or VSV-G pseudovirus. % Relative Luciferase Units (RLU) were normalized to the mean of the no-tetracycline controls. n = 12 wells from 4 experiments, two-way ANOVA with multiple comparisons. B) PAC inducible HEK 293T ACE2 cells were infected with pseudoviruses carrying different SARS-CoV-2 spike proteins. SARS-CoV-2 XBB.1.16 is labelled 1.16 in figure. n = 12 wells from 4 experiments, unpaired two-tailed student’s t- test with Welch’s correction. The % RLU between control and tetracycline induced conditions were statistically significant with p <0.0001 for all groups except VSV-G pseudoviruses. Bars represent mean ± SEM; ns = not significant, **** p < 0.0001 .

Article Snippet: To generate VSV-G pseudoviral particles, we transfected a 15 cm dish of HEK 293T cells with 3.5 μg of VSV-G along with 3.5 μg of packaging plasmids pMDL and pRSV, and 16 μg of luciferase construct (pHIV-Luc-ZsGreen, Addgene 39196).

Techniques: Luciferase, Infection, Two Tailed Test, Control

Journal: bioRxiv

Article Title: The proton-activated chloride channel inhibits SARS-CoV-2 spike protein-mediated viral entry through the endosomal pathway

doi: 10.1101/2025.03.12.642872

Figure Lengend Snippet: A) Endosomal pH measurement using ratiometric transferrin assay for control HEK 293T ACE2 cells and the inducible cells expressing PAC, Y10A/L13A, and L309C mutants. n = 3, one-Way ANOVA with multiple comparisons. B) SARS-CoV-2 spike-mediated pseudoviral entry for cells with or without expression of WT PAC, Y10A/L13A, and L309C mutants. % Relative Luciferase Units (RLU) were normalized to the mean of the no-tetracycline controls. n = 12 wells from 4 experiments, two-way ANOVA with multiple comparisons. C) Cathepsin inhibitor E64-d had no major additive effect on SARS-CoV-2 spike-mediated pseudoviral entry with PAC overexpression. % RLUs were normalized to the mean of the non-treated control. n = 15 wells from 5 experiments, one Way ANOVA with multiple comparisons. Bars represent mean ± SEM; ns = not significant, * p<0.05, **** p < 0.0001 .

Article Snippet: To generate VSV-G pseudoviral particles, we transfected a 15 cm dish of HEK 293T cells with 3.5 μg of VSV-G along with 3.5 μg of packaging plasmids pMDL and pRSV, and 16 μg of luciferase construct (pHIV-Luc-ZsGreen, Addgene 39196).

Techniques: Control, Expressing, Luciferase, Over Expression

Journal: bioRxiv

Article Title: The proton-activated chloride channel inhibits SARS-CoV-2 spike protein-mediated viral entry through the endosomal pathway

doi: 10.1101/2025.03.12.642872

Figure Lengend Snippet: A) Western blot validating the inducible expression of PAC and stable expression of ACE2 and TMPRSS2. Representative image of three experiments. B) SARS-CoV-2 spike-mediated pseudoviral entry for HEK 292T ACE2 cells with or without PAC and TMPRSS2 expression. % Relative Luciferase Units (RLU) were normalized to the mean of the controls that did not express PAC or TMPRSS2. n = 15 wells from 5 experiments, unpaired two-tailed student’s t- test with Welch’s correction. Bars represent mean ± SEM; **** p<0.0001 .

Article Snippet: To generate VSV-G pseudoviral particles, we transfected a 15 cm dish of HEK 293T cells with 3.5 μg of VSV-G along with 3.5 μg of packaging plasmids pMDL and pRSV, and 16 μg of luciferase construct (pHIV-Luc-ZsGreen, Addgene 39196).

Techniques: Western Blot, Expressing, Luciferase, Two Tailed Test