Journal: Nature Communications

Article Title: Targeted degradation of cell surface proteins through endocytosis triggered by cell-penetrating peptide-small molecule conjugates

doi: 10.1038/s41467-025-62776-w

Figure Lengend Snippet: a , b Western blot analysis of PD-L1 levels in MDA-MB-231 cells treated with BMS-CPP1 ( a ) or BMS-CPP2 ( b ) at indicated concentration, or indicated time. c , d Western blot analysis of PD-L1 levels in MDA-MB-231 cells treated with PD-LYSO ( c , d ) or BMS-CPP1 ( d ) for 8 h at indicated concentration. e Western blot analysis of HA-PD-L1 levels in HeLa cells (stably expressing HA-PD-L1) treated with BMS-CPP1 or BMS-CPP2 for 8 h at indicated concentration. f , g Immunofluorescence analysis of PD-L1 (red) degradation on cell membrane ( f ) or in whole cells ( g ). HeLa cells stably expressing PD-L1 were treated with BMS-CPP1, BMS-8, CPP1, or a combination of BMS-8 and CPP1. The nuclei were labeled by DAPI (blue). Scale bar, 10 μm. h Western blot analysis of PD-L1 levels in different cell lines treated with 25 nM of BMS-CPP1, BMS-8 or CPP1 for 8 h. i , j Western blot analysis of PD-L1 levels in MDA-MB-231 cells treated with 25 nM of BMS-CPP1 ( i ) or 5 nM of BMS-CPP2 ( j ) for 8 h along with bafilomycin A1 (100 μM) or MG132 (5 μM). k , l Western blot analysis of the inhibitory effect of Nystatin (50 μM), 7-KC (7-keto-cholesterol, 30 μM), CPZ (chlorpromazine, 10 μg/mL) or EIPA (ethylisopropylamiloride, 10 μg/mL) on the degradation of PD-L1 mediated by BMS-CPP1 ( k ) or BMS-CPP2 ( l ). Source data are provided as a Source Data file.

Article Snippet: The membranes were respectively incubated overnight with primary antibodies [PD-L1 antibody, Cell Signaling Technology (CST), rabbit, 1:1000; PD-L1 antibody, Proteintech, mouse, 1:5000; CA9 antibody, Cell Signaling Technology (CST), rabbit, 1:1000; CB 2 R antibody, Abcam, rabbit, 1:200; GAPDH antibody, Proteintech, mouse, 1:10000) at 4 o C with slightly shaking.

Techniques: Western Blot, Concentration Assay, Stable Transfection, Expressing, Immunofluorescence, Membrane, Labeling

Journal: Nature Communications

Article Title: Targeted degradation of cell surface proteins through endocytosis triggered by cell-penetrating peptide-small molecule conjugates

doi: 10.1038/s41467-025-62776-w

Figure Lengend Snippet: a Schematic illustration of the tumor-inhibition study and the general treatment procedure. Created in BioRender. OU, ZI. (2025) https://BioRender.com/8m9xnsp . b Body weight change curves of mice from day 6 to day 18. Data represent the mean ± SEM ( n = 5 mice per group). c Tumor growth curves for each group. Comparison of the tumor size after 18 days of different treatments. Data represent the mean ± SEM ( n = 5 mice per group). The statistical significance was assessed using two-way ANOVA. d Photograph of the peeled-off tumors of all four groups. e Comparison of the tumor weight after tumor dissection. Data represent the mean ± SEM ( n = 5 mice per group). The statistical significance was assessed using two-tailed Student’s t -tests. f Metastasis of tumor in the spleens of each group. g Western blot analysis of PD-L1 in B16F10 tumor tissues. Densitometry was used to calculate protein levels, and data were normalized to control. Data represent the mean ± SEM ( n = 3 mice per group). Statistical significance was assessed using two-tailed Student’s t -tests. h Representative immunohistochemical staining of PD-L1 in tumor tissues. Scale bar, 50 μm. Source data are provided as a Source Data file.

Article Snippet: The membranes were respectively incubated overnight with primary antibodies [PD-L1 antibody, Cell Signaling Technology (CST), rabbit, 1:1000; PD-L1 antibody, Proteintech, mouse, 1:5000; CA9 antibody, Cell Signaling Technology (CST), rabbit, 1:1000; CB 2 R antibody, Abcam, rabbit, 1:200; GAPDH antibody, Proteintech, mouse, 1:10000) at 4 o C with slightly shaking.

Techniques: Inhibition, Comparison, Dissection, Two Tailed Test, Western Blot, Control, Immunohistochemical staining, Staining

Journal: Nature Communications

Article Title: Targeted degradation of cell surface proteins through endocytosis triggered by cell-penetrating peptide-small molecule conjugates

doi: 10.1038/s41467-025-62776-w

Figure Lengend Snippet: a , b Western blot analysis of PD-L1 levels in MDA-MB-231 cells ( a ) or 4T1 cells with low expression level of αvβ3 integrin ( b ) treated with BMS-CPP1 or BMS-RGD for 8 h at indicated concentration. c , d Western blot analysis of PD-L1 levels in MDA-MB-231 cells treated with BMS-RGD ( c ) or BMS-CPP1 ( d ) for 8 h at indicated concentration. Source data are provided as a Source Data file.

Article Snippet: The membranes were respectively incubated overnight with primary antibodies [PD-L1 antibody, Cell Signaling Technology (CST), rabbit, 1:1000; PD-L1 antibody, Proteintech, mouse, 1:5000; CA9 antibody, Cell Signaling Technology (CST), rabbit, 1:1000; CB 2 R antibody, Abcam, rabbit, 1:200; GAPDH antibody, Proteintech, mouse, 1:10000) at 4 o C with slightly shaking.

Techniques: Western Blot, Expressing, Concentration Assay

Journal: Cell Communication and Signaling : CCS

Article Title: High expression of AMAP1, an ARF6 effector, is associated with elevated levels of PD-L1 and fibrosis of pancreatic cancer

doi: 10.1186/s12964-020-00608-8

Figure Lengend Snippet: High AMAP1 expression levels statistically correlate with PD-L1 expression in human PDACs and KPC tumors. a Representative IHC images of AMAP1 with IHC scores 0 to 3 and positive and negative staining of PD-L1 in clinical specimens, and their comparison with AMAP1 IHC scores. b IHC images and quantification of the PD-L1 staining of tumors formed by control (Irr) or AMAP1 -silenced ( shAMAP1 #1 and #2) KPC cells in C57BL/6 mice. Error bars represent the mean ± s.e.m. ** P < 0.01. c PD-L1 cell surface expression in IFNγ-treated or non-treated KPC cells, pretreated with shRNAs. MFI, median fluorescence intensity. Error bars represent the mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001. d Kaplan-Meier plots of the overall survival of patients with regard to PD-L1 positivity. P -values were obtained by t -tests ( a , b and c ) and by the log-rank test ( d ). Bars = 100 μm ( a and b )

Article Snippet: Other antibodies were purchased from commercial sources, as follows: rabbit monoclonal antibodies against PD-L1 (Cell Signaling) and phospho-FAK (Thermo Fisher Scientific), rabbit polyclonal antibody against PD-L1 (Novus), rabbit polyclonal antibody against collagen I (Proteintech), rabbit polyclonal antibody against FAK [ ], mouse monoclonal antibody against β-actin (Sigma-Aldrich).

Techniques: Expressing, Negative Staining, Comparison, Staining, Control, Fluorescence