Journal: EMBO Molecular Medicine

Article Title: GP64-pseudotyped lentiviral vectors target liver endothelial cells and correct hemophilia A mice

doi: 10.1038/s44321-024-00072-8

Figure Lengend Snippet: ( A , B ) Mean and SEM with single values of the VCN ( A ) or GFP-positive tissue ( B ) measured in the total liver of 8-week-old mice 1 week after i.v. administration of VSV.G-LV or GP64-LV at the indicated doses ( n = 5 mice per group). Mann–Whitney test. ( C ) Mean and SEM of the distribution of the GFP-positive tissue among the major liver cell types of mice shown in ( A , B ) at the indicated doses (KC Kupffer cells, LSEC liver sinusoidal endothelial cells, Hep hepatocytes). ( D ) Representative images and merge of immunofluorescent staining of liver analyzed in B and C at 4e10 TU/kg ( n = 5 mice). Red: F4/80-positive cells (KC); white: Lyve1-positive cells (LSEC); green: GFP (transduced cells); blue: Hoechst (nuclei). Orange arrows indicate GFP-positive LSEC. Blue arrows indicate GFP-positive KC. Scale bar: 250 µm. .

Article Snippet: The nPC were subsequently incubated with the following monoclonal antibodies: e-fluor 450-conjugated anti-CD45 (30-F11, e-Bioscience), Allophycocyanin (APC)-conjugated anti-CD31 (MEC13.3, BD Biosciences), phycoerythrin (PE)-conjugated F4/80 (CI:A3-1, Biorad), PE-Cy5-conjugated anti-CD45R/B220 (from BD Biosciences), PE-Cy7-conjugated anti-CD11c (N418, e-Bioscience). nPC subpopulations (LSEC, KC, pDC) were sorted by FACS, BD FACSAria™ Fusion Cell Sorter (BD Biosciences); the nPC contaminating the PC suspension, were removed by FACS excluding cells labeled by APC-conjugated anti-CD31/anti-CD45 cocktail, thus obtaining sorted hepatocytes (Hep) using BD FACSMelody™ Cell Sorter (BD Biosciences) or MoFlo Astrios EQ Cell Sorter (Beckman Coulter).

Techniques: MANN-WHITNEY, Staining

Journal: EMBO Molecular Medicine

Article Title: GP64-pseudotyped lentiviral vectors target liver endothelial cells and correct hemophilia A mice

doi: 10.1038/s44321-024-00072-8

Figure Lengend Snippet: ( A – C ) Single values, mean and SEM of the % of KC ( A ), hepatocytes ( B ) or LSEC ( C ) in GFP-positive cells shown in Fig. , reported here separately for statistical analysis. n = 5 per group. Mann–Whitney test. ( D ). Representative merge images of immunofluorescent staining of liver sections analyzed in Fig. at the indicated doses. Red: F4/80-positive cells (KC); White: Lyve1-positive cells (LSEC); Green: GFP (transduced cells); Blue: Hoechst (nuclei). Scale bar: 250 µm. Images of the 4e10 TU/kg LV dose are also presented in Fig. . ( E ) Representative images of immunofluorescent staining of a liver section from an untreated control mouse. Red: F4/80-positive cells (KC); White: Lyve1-positive cells (LSEC); Green: GFP (transduced cells); Blue: Hoechst (nuclei). Scale bar: 250 µm.

Article Snippet: The nPC were subsequently incubated with the following monoclonal antibodies: e-fluor 450-conjugated anti-CD45 (30-F11, e-Bioscience), Allophycocyanin (APC)-conjugated anti-CD31 (MEC13.3, BD Biosciences), phycoerythrin (PE)-conjugated F4/80 (CI:A3-1, Biorad), PE-Cy5-conjugated anti-CD45R/B220 (from BD Biosciences), PE-Cy7-conjugated anti-CD11c (N418, e-Bioscience). nPC subpopulations (LSEC, KC, pDC) were sorted by FACS, BD FACSAria™ Fusion Cell Sorter (BD Biosciences); the nPC contaminating the PC suspension, were removed by FACS excluding cells labeled by APC-conjugated anti-CD31/anti-CD45 cocktail, thus obtaining sorted hepatocytes (Hep) using BD FACSMelody™ Cell Sorter (BD Biosciences) or MoFlo Astrios EQ Cell Sorter (Beckman Coulter).

Techniques: MANN-WHITNEY, Staining