Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Aug 22;15:7237. doi: 10.1038/s41467-024-51720-z

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

PMC Copyright notice

Fig. 3 — a Schematic illustration of live-cell confocal microscopy assay. b Live-cell images of B16F10 cells treated with ^FITCαPD-L1 (3.3 nM) or ^FITCαPD-L1-NP (^FITCPD-L1-equiv., 3.3 nM) for 4 h. Scale bar, 10 μm. The images are representative of n = 3 biological replicates. c, d Western blot analysis of total (c) and membrane-associated (d) PD-L1 in B16F10 cells receiving different treatments for 24 h. αPD-L1-equiv. concentration, 3.3 nM; NP-equiv. concentration, 25 μg mL⁻¹. The blots are representative of n = 3 biological replicates. e IF of surface PD-L1 in B16F10 cells treated with αPD-L1 (3.3 nM) or αPD-L1-NP (PD-L1-equiv., 3.3 nM) for 24 h. Scale bar, 20 μm. The IF images are representative of n = 3 biological replicates. f Western blot analysis of B16F10 cells treated with αPD-L1-NP (3.3 nM) for 12 or 24 h in the presence or absence of 0.1 mg mL⁻¹ leupeptin (LPT). The blots are representative of n = 3 biological replicates. g–j Western blot analysis of PD-L1 in B16F10 cells treated with αPD-L1-NP at the indicated concentrations for 24 h (g and h) or at 3.3 nM for the indicated durations (i and j) The blots are representative of n = 3 biological replicates. k–o In vivo antitumor study of αPD-L1-NP in B16F10 tumor-bearing C57BL/6 mice. Mice (n = 5 mice per group) were treated intratumorally (i.t.) with PBS, αPD-L1 (2.0 mg kg⁻¹), or αPD-L1-NP (αPD-L1-equiv. dose, 2.0 mg kg⁻¹) for three times, respectively. k Schematic diagram outlining the experimental design. l Tumor growth curves of mice receiving different treatments. m Image of tumors resected after animal euthanasia. n Immunofluorescence staining of PD-L1 in tumor sections. Scale bar, 100 μm. o Western blot analysis of PD-L1 levels in tumor lysates. The blots are representative of n = 3 biological replicates. All the uncropped blots are included in the Source Data file. Data are presented as mean ± SD. Statistical significance was calculated via one-way ANOVA with Dunnett’s post hoc test. **p < 0.01; ***p < 0.001. Source data are provided as a Source Data file.