Journal: iScience

Article Title: Enhanced reverse zoonotic potential and immune evasion by omicron JN.1 variant

doi: 10.1016/j.isci.2025.112824

Figure Lengend Snippet: Transduction of various omicron S pseudovirions on 293/hACE2 and Calu3 cells (A) S proteins incorporation in pseudovirions. Pseudovirions with different omicron S proteins were pelleted down by centrifugation through 20% sucrose cushion and separated in a 10% SDS-PAGE. Detection of S proteins in pseudovirions was performed by Western blot using rabbit polyclonal anti-S2 antibodies. The p24 served as the loading controls. Experiments were done three times and one representative was shown. (B and C) Entry of pseudovirions of omicron variants. HEK 293/hACE2 and Calu3 cells were transduced with omicron S pseudovirions and lysed at 40 h post-transduction. The transduction efficiencies were determined according to luciferase activities. Experiments were done three times in triplicate and one representative is shown. The statistical difference relative to WT was determined by one-way ANOVA with Dunnett’s multiple testing correction ( n = 3). The statistical difference between BA.2.86 and JN.1 was determined by a two-tailed t-test ( n = 3). Error bars indicate SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Rabbit polyclonal against HIV-1 Gag-p24 antibodies , SinoBiological Inc (Beijing, China) , Cat#11695-T62.

Techniques: Transduction, Centrifugation, SDS Page, Western Blot, Luciferase, Two Tailed Test

Journal: Cell Discovery

Article Title: A potent and broad CD4 binding site neutralizing antibody with strong ADCC activity from a Chinese HIV-1 elite neutralizer

doi: 10.1038/s41421-025-00808-x

Figure Lengend Snippet: a , b Neutralization potency ( a ) and breadth ( b ) of FD22, in comparison with VRC01, against a 145-isolate Env-pseudovirus panel. c , d The neutralization activity ( c ) and percentage ( d ) of FD22 were evaluated against different clades of HIV-1 pseudoviruses from a 145-virus panel, including the two predominant circulating strains in China, CRF01_AE and CRF07_BC. VRC01 served as a control. e Activation of HIV-1 latency in ACH-2 cells. HIV-1 latency reversal in ACH-2 cells was induced by treatment with TNF-α or panobinostat for 48 h. Viral reactivation was assessed by quantifying p24 antigen concentrations using a standardized ELISA. f Inhibitory activity of FD22 against latent HIV-1 in ACH-2 cells activated by TNF-α or panobinostat, as measured in TZM-bl cells. ACH-2 cells were treated with TNF-α (10 ng/mL) or panobinostat (1 μM) to induce HIV-1 reactivation. After activation, FD22 was added at various concentrations to assess its inhibitory activity. Reactivated virus production was measured by coculturing supernatants from treated ACH-2 cells with TZM-bl indicator cells.

Article Snippet: Rabbit polyclonal anti-Gag-p24 antibodies (Sino Biological Inc.) were used to coat ELISA plates.

Techniques: Neutralization, Comparison, Activity Assay, Virus, Control, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Packaging cells for lentiviral vectors generated using the cumate and coumermycin gene induction systems and nanowell single-cell cloning

doi: 10.1016/j.omtm.2023.02.013

Figure Lengend Snippet: Regulation of expression of LV gene components in the packaging cells (A) Expression of VSV-G, Rev, and Gag by the packaging cells (clone 3D4) was analyzed by western blots after induction with cumate and coumermycin. The same amount of total protein from the cell lysate was used for the analysis. Cells were harvested before (0) or after 24, 48, and 72 h of induction in the absence or presence of sodium butyrate (+B) (added 18 h post induction). 293SF-CymR/λR-GyrB cells were used as a negative control (CT). The position of molecular weight marker in kDa (MW) is indicated. The position of VSV-G, Rev, the Gag polyprotein (GagPP), and p24 is indicated. Note the presence of non-specific bands (∗) in the negative control when using the anti-VSV-G antibody. (B) Analysis of the induction level for VSV-G, GagPP, and Gag p24. The western signal before and at 48 h post induction in the absence or presence of sodium butyrate was measured using a digital imaging system. The values were normalized to the signal of β-actin obtained after stripping and incubating the membrane with an anti-actin antibody. The experiment was done in triplicate (N = 3). Value are the means ± SD. Means significantly different from each other: ∗∗p < 0.01, ∗p < 0.05.

Article Snippet: The same amount of total protein (40 μg) was separated through a NuPAGE 4–12% Bis-Tris Gel, (Invitrogen, Carlsbad, CA) and analyzed by western blotting using a rabbit polyclonal HIV p24 Ab (ProSci, Poway CA, catalog no. 7313), a rabbit polyclonal anti-VSV-G tag antibody (ab83196, abcam, Waltham MA), or anti-HIV1 Rev mouse monoclonal antibody (ab85529, abcam), followed by a horseradish peroxidase-conjugated donkey anti-rabbit immunoglobulin (Ig) G antibody or a horseradish peroxidase-conjugated sheep anti-mouse IgG antibody (GE Healthcare, Buckinghamshire, UK).

Techniques: Expressing, Western Blot, Negative Control, Molecular Weight, Marker, Imaging, Stripping Membranes, Membrane