Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Serum factors create species-specific barriers to hepatic gene transfer by lipid nanoparticles in liver-humanized mice

doi: 10.1016/j.omtm.2025.101470

Figure Lengend Snippet: Comparative histopathology and putative LNP receptor expression in chimeric NSG-PiZ and FRG mouse livers Livers from liver-humanized NSG-PiZ or FRG mice at 24- or 48-h post treatment with PBS, or LNP1–3 formulated with DasherGFP mRNA (A and B). Paraffin-embedded sections from representative livers were stained with H&E (A). Scale bar, 100 μm. Frozen sections from representative livers were stained with Oil red O and counterstained with hematoxylin (B). Scale bars, 1 mm (inset image) and 100 μm (main image). Dashed lines indicate approximate borders between human and mouse hepatocytes. H, human hepatocytes; M, mouse hepatocytes. snRNA-seq and scRNA-seq analysis of putative LNP receptor mRNA levels in hepatocytes of chimeric FRG and NSG-PiZ mice (C). (D) Detection of ALB , A PO E , LDLR , ASGR1 , and ASGR2 mRNA in human and mouse hepatocytes from NSG-PiZ and FRG mice. (E) The percentage of total reads per cell for ALB , APOE , LDLR , ASGR1 , and ASGR2 in human (H) or mouse (M) hepatocytes determined by scRNA-seq (NSG-PiZ) or snRNA-seq (FRG). For the scRNA-seq analysis, cells were pooled from four low engrafted chimeric NSG-PiZ mice prior to single-cell analysis. For the snRNA-seq analysis, reads were pooled from two highly engrafted chimeric FRG mice or two low engrafted chimeric FRG mice from a previously published dataset (Bioproject: PRJNA857105; GEO: GSM6319631 , GSM6319632 , GSM6319637 , GSM6319638 ; Cabanes-Creus et al. ). FRG reads were processed as previously described (Cabanes-Creus et al. ) and scRNA-seq reads were processed as described in the methods. (C) was created using Biorender.

Article Snippet: Serum from chimeric NSG-PiZ mice (HuAlb >1 mg/mL) was depleted of ApoE via immunoprecipitation using a rabbit anti-human ApoE polyclonal antibody (cat # VPA00665 , Bio-Rad, Hercules, CA) and protein G spin trap columns (Cat #28903134, Marlborough, MA) according to the manufacturer’s instructions.

Techniques: Histopathology, Expressing, Staining, Single-cell Analysis

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Serum factors create species-specific barriers to hepatic gene transfer by lipid nanoparticles in liver-humanized mice

doi: 10.1016/j.omtm.2025.101470

Figure Lengend Snippet: NSG-PiZ serum factors impact LNP transfection of human hepatocytes (A) Flow cytometry and immunofluorescence analysis of GFP expression in mouse and human (HLA+ or hCK18+) hepatic cells from chimeric NSG-PiZ mice 24 h after intravenous delivery of SM-102 LNPs (1.3 mg/kg). Chimeric NSG-PiZ mice were administered PBS ( n = 2) or SM-102 LNPs ( n = 4). Representative scatterplots and histograms show relative GFP expression in human or mouse cells for two PBS- and three SM-102 LNP-treated mice along with mouse HuAlb concentrations prior to LNP administration. Representative immunofluorescence staining of paraffin sections for the fourth SM-102 LNP-treated mouse with antibodies specific for GFP (green) and hCK18 (red). (B) serum impacts transfection of PHH in vitro . SM-102 LNP was preincubated with the indicated serum for 15 min at room temperature (RT) prior to addition to media. GFP expression was imaged 6 h after after LNP administration. (C) Inhibition of PHH transfection by chimeric NSG-PiZ mouse serum. SM-102 LNPs, LP01 LNPs, or ALC0315 LNPs were pre-treated with normal mouse serum for 15 min at RT prior to addition to media containing 2% chimeric serum. GFP expression was imaged 6 h after after LNP administration. (D) Inhibition of PHH transfection by recombinant human ApoE. SM-102 LNP was pre-treated with normal mouse serum for 15 min at RT prior to addition to media containing recombinant human ApoE at the indicated concentration. (E) Inhibition of PHH transfection by ApoE depleted chimeric NSG-PiZ mouse serum. Western blot for ApoE using chimeric NSG-PiZ serum, anti-ApoE depleted chimeric NSG-PiZ serum, or control anti-Flag depleted chimeric NSG-PiZ serum (left). SM-102 LNP was pre-treated with normal mouse serum for 15 min at RT prior to addition to media containing 2% chimeric NSG-PiZ serum deleted of ApoE. GFP expression was imaged 6 h after after LNP administration. (F) Inhibition of PHH transfection by naive NSG-PiZ mouse serum. SM-102 LNP was pre-treated with normal mouse serum for 15 min at RT prior to addition to media containing 2% chimeric or naive NSG-PiZ serum. GFP expression was imaged 6 h after after LNP administration. (G) NSG mouse serum supports SM-102 transfection of PHH. SM-102 LNP was pre-treated with NSG serum or chimeric NSG-PiZ serum for 15 min at RT prior to addition to media. GFP expression was imaged 6 h after LNP administration. The experiments shown in (C–F) were performed concurrently and for comparison, the same representative image for SM-102 transfection of PHH after pre-incubation with normal mouse serum is shown as the upper left image within each panel.

Article Snippet: Serum from chimeric NSG-PiZ mice (HuAlb >1 mg/mL) was depleted of ApoE via immunoprecipitation using a rabbit anti-human ApoE polyclonal antibody (cat # VPA00665 , Bio-Rad, Hercules, CA) and protein G spin trap columns (Cat #28903134, Marlborough, MA) according to the manufacturer’s instructions.

Techniques: Transfection, Flow Cytometry, Immunofluorescence, Expressing, Staining, In Vitro, Inhibition, Recombinant, Concentration Assay, Western Blot, Control, Comparison, Incubation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Enhancement of keratinocyte growth factor potential in inducing adipose‐derived stem cells differentiation into keratinocytes by collagen‐targeting

doi: 10.1111/jcmm.17619

Figure Lengend Snippet: Western blot analysis of purified KGF and vibrio CBD‐KGF. Lane M: Protein marker; lane 1: purified KGF; lane 2: purified vibrio CBD‐KGF. Western blot analysis was carried out using rabbit anti‐KGF antibody and goat anti‐rabbit HRP conjugated antibody.

Article Snippet: Serial dilutions of vibrio CBD‐KGF and KGF (25, 50, 100, 150, and 200 ng/ml) were applied to the plate, followed by incubation at 37°C for 2 h. Wells were washed extensively with PBS‐T, and after adding rabbit anti‐KGF antibody (1:1000, Sigma, USA), the plate was kept at 4°C overnight.

Techniques: Western Blot, Purification, Marker

Journal: Journal of Cellular and Molecular Medicine

Article Title: Enhancement of keratinocyte growth factor potential in inducing adipose‐derived stem cells differentiation into keratinocytes by collagen‐targeting

doi: 10.1111/jcmm.17619

Figure Lengend Snippet: Western blot analysis of purified KGF and vibrio CBD‐KGF. Lane M: Protein marker; lane 1: purified KGF; lane 2: purified vibrio CBD‐KGF. Western blot analysis was carried out using rabbit anti‐KGF antibody and goat anti‐rabbit HRP conjugated antibody.

Article Snippet: The following day, the membrane was incubated with rabbit anti‐KGF antibody (Sigma, USA), diluted (1:1000) in the blocking buffer, for 2 h at room temperature by gentle agitation.

Techniques: Western Blot, Purification, Marker

Journal: Breast cancer research and treatment

Article Title: Breast Cancer, Diabetes Mellitus and Glucagon-Like Peptide-1 Receptor Toward Exploring Their Possible Associations.

doi: 10.1007/s10549-021-06288-3

Figure Lengend Snippet: Fig. 1 Representative illustra- tion of immunochemical find- ings in breast carcinoma cases examined in this study. 0, 1+, 2+ and 3+ correspond to the intensity score of immunoreac- tivities obtained. The positive controls were pancreatic islets of Langerhans with positive expression of GLP-1R (a), blad- der cancer tumor with that of FGF7 (b) and lung cancer tumor with that of FGFR2 (c)

Article Snippet: A mouse monoclonal antibody against GLP-1R (ab166987, abcam, Cambridge UK), a rabbit polyclonal antibody against FGF7 (HPA043605, Atlas Antibodies, Stockholm Sweden) and a mouse monoclonal antibody against FGFR2 (ab58201, abcam, Cambridge UK) were used as primary antibodies in this study as summarized in Table 2.

Techniques: Expressing