Journal: Nature Communications

Article Title: Enhancing the potency of in vivo lentiviral vector mediated gene therapy to hepatocytes

doi: 10.1038/s41467-025-60073-0

Figure Lengend Snippet: a Mean with SEM of hFVIII concentration in the plasma of immune tolerant hemophilia A mice, treated with (red squares) or without (black squares) anti-IFNAR1 Ab (50 mg/kg, i.v.) or with a control Ab (immunoglobulin G control, IgG control, pink squares) 3 h before LV-FVIII (4–7 × 10 10 TU/kg). Pool of two independent experiments. Kruskal-Wallis test with Dunn’s multiple comparisons test over LV-FVIII control group, performed at the last time-point. b Single values and mean with SEM of VCN measured in the indicated fractionated or FACS-sorted liver cell types of mice in ( a ) 5 months post-LV. Two-tailed Mann-Whitney test. c Mean with SEM of hFIX concentration in the plasma of WT mice, treated with Bortezomib (1 mg/kg, blue dots) and/or Anti-IFNAR1 Ab (50 mg/kg, purple or red dots, as indicated) or saline (black dots), 1 h or 3 h before LV-FIX (2.75 × 10 10 TU/kg). Two-tailed linear-mixed effects (LME) model (for complete analyses see Supplementary Table ) over LV-FIX group. d Single values and mean with SEM of VCN measured in fractionated and FACS-sorted liver cell types of mice in ( c ) 3 months post-LV. Kruskal-Wallis test with Dunn’s multiple comparisons test over LV-FIX control group. e Single values and mean with SEM of the percentage of GFP-positive hepatocytes (7 fields/sample) transduced with PGK.GFP LV (multiplicity of infection (MOI): 2), in the presence (blue triangles) or absence (black triangles) of Bortezomib (100 nM) at the time of transduction, and analyzed 7 days after transduction. As a control for inhibition of GFP degradation, Bortezomib (100 nM) was added 2 days after LV administration (condition Bort control, gray triangles). f Single values and mean with SEM of VCN measured in the above-described experiment. Kruskal-Wallis test with Dunn’s multiple comparisons test. Source data are provided as a Source Data file.

Article Snippet: Microtiter plates were coated with anti-hFVIII binding Ab (Green Mountain Antibodies #GMA8016, 0.2 μg/well in 0.1 M carbonate buffer, pH 9.6) overnight at 4 °C and then blocked for 1 h at room temperature with blocking buffer (PBS 0.05% Tween-20, 1 M NaCl, 10% heat-inactivated horse serum, Gibco).

Techniques: Concentration Assay, Clinical Proteomics, Control, Two Tailed Test, MANN-WHITNEY, Saline, Transduction, Infection, Inhibition