Fig 1: MET inhibitors suppress the immunosuppressive capabilities of recombinant HGF. A, B Peripheral blood mononuclear cells (PBMCs) activated with phytohemagglutinin (PHA, 10 µg/ml) were cultured either alone (A) or in the presence of hepatocyte growth factor (HGF, 75 ng/mL) and/or MET inhibitors (crizotinib, SGX-523). The MTS proliferation assay was employed to quantify lymphocyte/PBMC proliferation at 72 h, while cell supernatants were collected at 24 h for interferon-gamma (IFNγ) quantification. C, D Activated PBMCs were stained with CFSE and treated for 96 h with HGF (75 ng/mL) alone or in combination with pembrolizumab (20 nm) and/or MET inhibitors (crizotinib, SGX-523). The stained cells were collected, probed with PE-Cy7-conjugated anti-CD3, BV421-conjugated anti-CD45, and APC-conjugated anti-CD8 antibodies, and analyzed using flow cytometry. Data are presented as mean ± SEM of at least four independent experiments. *p < 0.05 and ***p < 0.001
Fig 2: Fraction of HGF released over a 28-day time period during culture with cells. Asterisks (*) indicate a significant difference (p < 0.05).
Fig 3: Comparison of HGF release from coated versus uncoated nanoparticles. Asterisks (*) indicate a significant difference (p < 0.05).
Fig 4: HGF + EGF mRNA-LNP treatments augment PHH engraftment by increasing the survival and proliferation of donor cells in p21/NSG-PiZ mice.a PHH transplantation experimental timeline and treatment groups. A single IV dose of AAV8-TBG-p21 was injected one week prior to cell transplantation, while mRNA-LNP were administered IV 5 h before transplantation and twice weekly for 2 weeks. Cryopreserved PHHs were thawed and transplanted into male NSG-PiZ mice via intrasplenic injection of 106 cells/mouse. Four groups were used: control Luc mRNA-LNP only, AAV8-TBG-p21 only, HGF + EGF mRNA-LNP only, and the combined group with both p21 and HGF + EGF injections. Recipient mice were given an EdU injection and sacrificed 2 weeks post transplantation, serum was collected weekly, and liver tissue harvested for analysis. b Representative images of hALB/Ku80/EdU immunofluorescence stain on liver sections 2 weeks post transplantation. c Quantification of the number of hALB+ Ku80+ PHH clusters per 10x image. d Quantification of the number of hALB+ Ku80+ PHHs per cluster. e Quantification of the number of hALB+ Ku80+ EdU+ proliferating PHHs per cluster. f Quantification of co-stained hALB+ Ku80+ area per 10x image. For all histology quantification c–f at least three images were averaged per mouse, each image from a different liver lobe. P values were calculated by unpaired two-sided student’s t test. g Human serum albumin levels measured via ELISA over the course of 2-week transplantation experiments. P values were calculated by ordinary one-way ANOVA. For all panels: n = 5 mice per group except in p21 only group where n = 4 mice as specified in figure key, each dot represents one mouse, error bars = SEM, scale = 100 µm, ns P > 0.05, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. Yellow—control, green—p21 only, teal—HGF + EGF only, blue—p21 & HGF + EGF treated group. Source data are provided as a Source Data file. a Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.
Fig 5: Comparison of total fraction of HGF release over 14 days during culture in static and dynamic conditions.
Supplier Page from Abcam for Human HGF ELISA Kit (Hepatocyte Growth Factor)