cf r d systems  (R&D Systems)

Journal: Cell death & disease

Article Title: SIRPα blockade therapy potentiates immunotherapy by inhibiting PD-L1 + myeloid cells in hepatocellular carcinoma.

doi: 10.1038/s41419-025-07779-7

Figure Lengend Snippet: Fig. 4 CD47 blockade therapy suppressed tumor progression in the PDX model. A–C A PDX tumor model was constructed using NSG mice using human HCC tissues. Schematic diagram showing the construction of a PDX tumor model and huPBMC injection through the tail vein. Mice (n = 6 for each group) were treated with anti-CD47 antagonist TTI-621 (8 mg/kg, three times a week for 3–4 weeks, i.p.) or vehicle (A). The tumor images (scale bar, 1 cm) (B), tumor weight (C) in the PDX tumor model using HCC tissues from patient 1. D–G The survival and tumor sizes of the tumor-bearing mice were recorded from patients 1–4. H, I mIF analysis of the tumor tissues in PDX model was used to detect the TIMs (CD11b), PD-L1, CD8+ T cells, and cytotoxic function (GZMB). Scale bar, 50 μm. J, K Western blot and qRT-PCR analysis were used to detect the PD-L1 expression in TIMs of tumor tissues in the PDX model. TIMs were obtained through magnetic bead sorting. All data presented are shown as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant.

Article Snippet: Myeloid cells with or without SIRPA overexpression were treated with anti-SIRPα mAb (5 μg/ml) or anti-CD47 mAb (10 μg/ml) compared with the control, followed by incubation with mCD47-Fc (Cat#1866-CD-050, R&D Systems) for 12 h. Cells were collected for analysis after cocultivation.

Techniques: Construct, Injection, Western Blot, Quantitative RT-PCR, Expressing

Journal: Cell death & disease

Article Title: SIRPα blockade therapy potentiates immunotherapy by inhibiting PD-L1 + myeloid cells in hepatocellular carcinoma.

doi: 10.1038/s41419-025-07779-7

Figure Lengend Snippet: Fig. 6 SIRPα/CD47 axis promoted migration, PD-L1 expression of macrophages and HCC growth via upregulating PI3K/AKT signaling. A Schematic of the in vitro coculture experiments with PMA-treated THP-1 cells cocultured with Hepa1–6 or LPC-H12 cells to mimic liver cancer microenvironment treated with anti-CD47 mAb (10 μg/ml) or not. Macrophages were incubated with mCD47-Fc for 12 h before analysis. B Western blot analysis was used to detect the expression of indicated proteins in macrophages. C, D Transwell assay was used to detect the migration of macrophages towards the supernatants of the indicated groups. Scale bar, 100 μm. E Orthotopic HCC model using Hepa1–6 was established. Wild- type C57BL/6 mice (n = 6 for each group) were treated with anti-SIRPα mAb (100 μg/mouse, i.p.) and/or GSK690693 (30 mg/kg/day, i.p.) compared with the control group. Representative livers in each group were photographed at the endpoint (Scale bar, 1 cm). F, G Statistical analysis of tumor volume and tumor weight of the mice. All data presented are shown as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant.

Article Snippet: Myeloid cells with or without SIRPA overexpression were treated with anti-SIRPα mAb (5 μg/ml) or anti-CD47 mAb (10 μg/ml) compared with the control, followed by incubation with mCD47-Fc (Cat#1866-CD-050, R&D Systems) for 12 h. Cells were collected for analysis after cocultivation.

Techniques: Migration, Expressing, In Vitro, Incubation, Western Blot, Transwell Assay, Control