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ATCC breast tumor cell line 4t1

│ Construction and characterization of PLT-TACs. A , Fast fabrication process of PLT-TACs via microfluidic device with a herringbone structure chip. B , TEM images and DLS size distribution of activated platelets (PLTs). C , TEM images and DLS size distribution of engineered activated platelets with medium density of anti-PD-L1 and P1 (PLT-TAC m ). D , SEM images of PLTs, E , SEM images of PLT-TAC m , white squares indicate zoom-in regions, red squares indicate morphology features on the platelet surface. F , Confocal images of Cy3-, Cy5-and Dio-labeled PLT-TACs. G , FCM analysis and 3D confocal visualization for verifying the co-installation of anti-PD-L1 and P1 in PLT-TACs. H , SDS-PAGE analysis of protein expression profiles in activated platelets and PLT-TAC m . I , FCM analysis of key platelet membrane proteins, CD41, CD47 and CD62P, expressed on PLT-TACs. J , AFM images of topography, height and Young's modulus mapping of PLTs and PLT-TAC m . Scale bars, 5 μm (n = 5 technical replicates). K , CLSM visualization of the localization of anti-PD-L1 and P1 in <t>4T1</t> cells after internalization for 3h, anti-PD-L1 was labeled with Cy5, P1 was labeled with Cy3, nuclei were stained with Hoechst 33342, and lysosomes were stained with lysotracker Green DND26. Scale bar, 10 μm. L , Fluorescence intensity profiles along the line drawn in K (n = 3 biological replicates).

Breast Tumor Cell Line 4t1, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 6856 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "“Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation"

Article Title: “Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation

Journal: Bioactive Materials

doi: 10.1016/j.bioactmat.2026.01.011

Figure Legend Snippet: │ Construction and characterization of PLT-TACs. A , Fast fabrication process of PLT-TACs via microfluidic device with a herringbone structure chip. B , TEM images and DLS size distribution of activated platelets (PLTs). C , TEM images and DLS size distribution of engineered activated platelets with medium density of anti-PD-L1 and P1 (PLT-TAC m ). D , SEM images of PLTs, E , SEM images of PLT-TAC m , white squares indicate zoom-in regions, red squares indicate morphology features on the platelet surface. F , Confocal images of Cy3-, Cy5-and Dio-labeled PLT-TACs. G , FCM analysis and 3D confocal visualization for verifying the co-installation of anti-PD-L1 and P1 in PLT-TACs. H , SDS-PAGE analysis of protein expression profiles in activated platelets and PLT-TAC m . I , FCM analysis of key platelet membrane proteins, CD41, CD47 and CD62P, expressed on PLT-TACs. J , AFM images of topography, height and Young's modulus mapping of PLTs and PLT-TAC m . Scale bars, 5 μm (n = 5 technical replicates). K , CLSM visualization of the localization of anti-PD-L1 and P1 in 4T1 cells after internalization for 3h, anti-PD-L1 was labeled with Cy5, P1 was labeled with Cy3, nuclei were stained with Hoechst 33342, and lysosomes were stained with lysotracker Green DND26. Scale bar, 10 μm. L , Fluorescence intensity profiles along the line drawn in K (n = 3 biological replicates).

Techniques Used: Labeling, SDS Page, Expressing, Membrane, Staining, Fluorescence

│ Surface binding, internalization, and PD-L1 degradation. A , Schematic illustration of the trafficking route of PLT-TACs. B , Fluorescence intensity analysis and histogram plots of PD-L1 binding to 4T1 cell surface after treatment with free anti-PD-L1, PLT-anti-PD-L1 or different PLT-TACs for 30min at 4 °C, measured by FCM (n = 3 biologically independent samples). C , Fluorescence intensity analysis and histogram plots of internalized PLT-anti-PD-L1 or different PLT-TACs in IFN-γ pre-treated 4T1 cells for 1, 2, or 4 h at 37 °C, measured by FCM (n = 3 biologically independent samples). D , Visualization of internalization of PLT-anti-PD-L1 or different PLT-TACs in IFN-γ pre-treated 4T1 cells after treatment for 2 h, observed by 40 × CLSM. Scale bar, 10 μm. E , Redistribution of PLT-TAC m membrane after treatment for 2 h or 4 h, observed by oil 63 × CLSM. Platelets were stained with Dio. Scale bar, 5 and 20 μm. F , Visualization and fluorescence intensity profiles along the line of colocalization of PLT-anti-PD-L1 or different PLT-TACs with lysosomes in IFN-γ pre-treated 4T1 cells after treatment for 4 h, observed by oil 63 × CLSM. Lysosomes were stained with lysotracker Green DND26. Scale bar, 10 μm. G , Mean fluorescence intensity of levels of PD-L1 in IFN-γ pre-treated 4T1, measured by FCM after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). H , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). I , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m with different concentration (0, 0.025, 0.25, 1.25, 2.5, 5 μM) for 36 h (n = 3 biologically independent samples). J , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) with different time (0, 6, 12, 24, 36, 48 h) (n = 3 biologically independent samples). K , Immunofluorescence of surface PD-L1 in IFN-γ pre-treated 4T1 cells treated with PLT-TAC m (5 μM) for 36 h. Scale bar, 20 μm. L , Visualization of PD-L1 degradation in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) for 0 h or 24 h. Scale bar, 5 μm. M , Western blot analysis of CD24, CD71 and CD47 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) for 36 h (n = 3 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Figure Legend Snippet: │ Surface binding, internalization, and PD-L1 degradation. A , Schematic illustration of the trafficking route of PLT-TACs. B , Fluorescence intensity analysis and histogram plots of PD-L1 binding to 4T1 cell surface after treatment with free anti-PD-L1, PLT-anti-PD-L1 or different PLT-TACs for 30min at 4 °C, measured by FCM (n = 3 biologically independent samples). C , Fluorescence intensity analysis and histogram plots of internalized PLT-anti-PD-L1 or different PLT-TACs in IFN-γ pre-treated 4T1 cells for 1, 2, or 4 h at 37 °C, measured by FCM (n = 3 biologically independent samples). D , Visualization of internalization of PLT-anti-PD-L1 or different PLT-TACs in IFN-γ pre-treated 4T1 cells after treatment for 2 h, observed by 40 × CLSM. Scale bar, 10 μm. E , Redistribution of PLT-TAC m membrane after treatment for 2 h or 4 h, observed by oil 63 × CLSM. Platelets were stained with Dio. Scale bar, 5 and 20 μm. F , Visualization and fluorescence intensity profiles along the line of colocalization of PLT-anti-PD-L1 or different PLT-TACs with lysosomes in IFN-γ pre-treated 4T1 cells after treatment for 4 h, observed by oil 63 × CLSM. Lysosomes were stained with lysotracker Green DND26. Scale bar, 10 μm. G , Mean fluorescence intensity of levels of PD-L1 in IFN-γ pre-treated 4T1, measured by FCM after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). H , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). I , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m with different concentration (0, 0.025, 0.25, 1.25, 2.5, 5 μM) for 36 h (n = 3 biologically independent samples). J , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) with different time (0, 6, 12, 24, 36, 48 h) (n = 3 biologically independent samples). K , Immunofluorescence of surface PD-L1 in IFN-γ pre-treated 4T1 cells treated with PLT-TAC m (5 μM) for 36 h. Scale bar, 20 μm. L , Visualization of PD-L1 degradation in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) for 0 h or 24 h. Scale bar, 5 μm. M , Western blot analysis of CD24, CD71 and CD47 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) for 36 h (n = 3 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques Used: Binding Assay, Fluorescence, Membrane, Staining, Saline, Western Blot, Concentration Assay, Immunofluorescence

│ Internalization mechanism analysis of PLT-TACs. A, Mutated P1 peptide (M1) designed using ProteinMPNN. B , Fluorescence intensity analysis of internalized M1-modified PLT-TAC m (M1) for 1, 2, or 4 h at 37 °C, measured by FCM (n = 3 biologically independent samples). C , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 cells after treated with PLT-TAC m (M1) and PLT-TAC m (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). D , Dynamic internalization process of PLT-TAC m at different time points, observed by confocal microscopy. Scale bar = 5 μm. E , Visualization and fluorescence intensity profiles along the line of the colocalization of PLT-TACs with endo/lysosomes in IFN-γ pre-treated 4T1 cells after treated for 2 h or 4 h, observed by oil 63 × CLSM (Early endosomes were stained by EEA1, lysosomes were stained by LAMP1, anti-PD-L1 was labeled with Cy5, nuclei were stained with Hoechst 33342.). Scale bar, 10 μm (n = 3 biologically independent samples). F , Visualization of the amount of lysosome biogenesis in IFN-γ pre-treated 4T1 cells after treated with PLT-anti-PD-L1 or PLT-TACs for 2 h or 4 h, observed by oil 63 × CLSM. Lysosomes were stained by LAMP1. Scale bar, 10 μm. G , Quantification of LAMP1 positive puncta in saline and PLT-TAC m treated cells after 4 h treatment. H , Western blot analysis of LAMP1 after treated with saline and PLT-TAC m for 36 h. I , Visualization of the colocalization of PLT-anti-PD-L1 and PLT-TAC m with LC3-II-labeled autophagosomes (yellow box) in IFN-γ pre-treated 4T1 cells after treatment for 4 h, observed by oil 63 × CLSM. Autophagosomes were stained by LC3-Ⅱ, anti-PD-L1 was labeled with Cy5, nuclei were stained with Hoechst 33342. Scale bar, 10 μm. J , Western blot analysis of LC3-I and LC3-II in after treated with PLT-anti-PD-L1 and PLT-TAC m for 36 h. K , Visualization of localization of TFEB and statistical analysis of cells with nuclear localization of TFEB after treated with PLT-anti-PD-L1 and PLT-TAC m for 36 h. Scale bar, 20 μm. (n = 3 biologically independent samples). L , Fluorescence intensity of internalized PLT-anti-PD-L1 and PLT-TAC m in IFN-γ pre-treated 4T1 cells after treatment for 2 h in presence of chlorpromazine, nystatin, colchicine, and cytochalasin D, measured by FCM (n = 3 biologically independent samples). M , Visualization of internalization of PLT-TACs and lysosome biogenesis in IFN-γ pre-treated 4T1 cells after treatment for 2 h in presence of chlorpromazine, nystatin, colchicine, and cytochalasin D, observed by oil 63 × CLSM. Scale bar, 10 μm. N, SEM images of internalization of PLT-TAC m by IFN-γ pre-treated 4T1 cells. Scale bar, 10 μm. Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Figure Legend Snippet: │ Internalization mechanism analysis of PLT-TACs. A, Mutated P1 peptide (M1) designed using ProteinMPNN. B , Fluorescence intensity analysis of internalized M1-modified PLT-TAC m (M1) for 1, 2, or 4 h at 37 °C, measured by FCM (n = 3 biologically independent samples). C , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 cells after treated with PLT-TAC m (M1) and PLT-TAC m (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). D , Dynamic internalization process of PLT-TAC m at different time points, observed by confocal microscopy. Scale bar = 5 μm. E , Visualization and fluorescence intensity profiles along the line of the colocalization of PLT-TACs with endo/lysosomes in IFN-γ pre-treated 4T1 cells after treated for 2 h or 4 h, observed by oil 63 × CLSM (Early endosomes were stained by EEA1, lysosomes were stained by LAMP1, anti-PD-L1 was labeled with Cy5, nuclei were stained with Hoechst 33342.). Scale bar, 10 μm (n = 3 biologically independent samples). F , Visualization of the amount of lysosome biogenesis in IFN-γ pre-treated 4T1 cells after treated with PLT-anti-PD-L1 or PLT-TACs for 2 h or 4 h, observed by oil 63 × CLSM. Lysosomes were stained by LAMP1. Scale bar, 10 μm. G , Quantification of LAMP1 positive puncta in saline and PLT-TAC m treated cells after 4 h treatment. H , Western blot analysis of LAMP1 after treated with saline and PLT-TAC m for 36 h. I , Visualization of the colocalization of PLT-anti-PD-L1 and PLT-TAC m with LC3-II-labeled autophagosomes (yellow box) in IFN-γ pre-treated 4T1 cells after treatment for 4 h, observed by oil 63 × CLSM. Autophagosomes were stained by LC3-Ⅱ, anti-PD-L1 was labeled with Cy5, nuclei were stained with Hoechst 33342. Scale bar, 10 μm. J , Western blot analysis of LC3-I and LC3-II in after treated with PLT-anti-PD-L1 and PLT-TAC m for 36 h. K , Visualization of localization of TFEB and statistical analysis of cells with nuclear localization of TFEB after treated with PLT-anti-PD-L1 and PLT-TAC m for 36 h. Scale bar, 20 μm. (n = 3 biologically independent samples). L , Fluorescence intensity of internalized PLT-anti-PD-L1 and PLT-TAC m in IFN-γ pre-treated 4T1 cells after treatment for 2 h in presence of chlorpromazine, nystatin, colchicine, and cytochalasin D, measured by FCM (n = 3 biologically independent samples). M , Visualization of internalization of PLT-TACs and lysosome biogenesis in IFN-γ pre-treated 4T1 cells after treatment for 2 h in presence of chlorpromazine, nystatin, colchicine, and cytochalasin D, observed by oil 63 × CLSM. Scale bar, 10 μm. N, SEM images of internalization of PLT-TAC m by IFN-γ pre-treated 4T1 cells. Scale bar, 10 μm. Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques Used: Fluorescence, Modification, Western Blot, Confocal Microscopy, Staining, Labeling, Saline

│ Efficacy and versatility of PLT-TACs. A , Molecular mechanism underlying PD-L1 degradation-induced apoptosis. B , C , FCM analysis and quantification of cell apoptosis after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h, followed by annexin V-FITC/PI staining (n = 3 biologically independent samples). D , E , Images and statistical analysis of wound healing assay in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h. Scale bar, 200 μm (n = 3 biologically independent samples). F , G , Images and statistical analysis of transwell cell invasion assay in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h. Scale bar, 200 μm (n = 3 biologically independent samples). H , Structures of CD71 and T7 peptide and the binding site of CD71/T7 complex, calculated by AlphaFold 3. I , Construction and characterization of PLT-TAC T7 by TEM and confocal microscopy. J , Mean fluorescence intensity of levels of surface CD71 in 4T1 cells after treated with saline, PLT-anti-PD-L1, or different PLT-TAC T7 (at an equivalent amount of 10 μM) for 36 h, measured by FCM (n = 3 biologically independent samples). K , Western blot analysis of CD71 in IFN-γ pre-treated 4T1 cells after treated with saline, and different PLT-TAC T7 (at an equivalent amount of 10 μM) for 36 h (n = 3 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Figure Legend Snippet: │ Efficacy and versatility of PLT-TACs. A , Molecular mechanism underlying PD-L1 degradation-induced apoptosis. B , C , FCM analysis and quantification of cell apoptosis after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h, followed by annexin V-FITC/PI staining (n = 3 biologically independent samples). D , E , Images and statistical analysis of wound healing assay in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h. Scale bar, 200 μm (n = 3 biologically independent samples). F , G , Images and statistical analysis of transwell cell invasion assay in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h. Scale bar, 200 μm (n = 3 biologically independent samples). H , Structures of CD71 and T7 peptide and the binding site of CD71/T7 complex, calculated by AlphaFold 3. I , Construction and characterization of PLT-TAC T7 by TEM and confocal microscopy. J , Mean fluorescence intensity of levels of surface CD71 in 4T1 cells after treated with saline, PLT-anti-PD-L1, or different PLT-TAC T7 (at an equivalent amount of 10 μM) for 36 h, measured by FCM (n = 3 biologically independent samples). K , Western blot analysis of CD71 in IFN-γ pre-treated 4T1 cells after treated with saline, and different PLT-TAC T7 (at an equivalent amount of 10 μM) for 36 h (n = 3 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques Used: Saline, Staining, Wound Healing Assay, Invasion Assay, Binding Assay, Confocal Microscopy, Fluorescence, Western Blot

│ In vivo antitumor and degradation efficacy by PLT-TACs. A , Schematic schedule of the treatment and evaluation in BALB/c mice with 4T1 orthotopic tumor model. B , Tumor growth curves, C , images of dissected tumors, D , tumor weights, E , survival curves, F , body weight changes, G , tumor growth kinetics from the control and treatment groups (saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h ) at a dosage of 5 mg/kg (n = 6 biologically independent samples). H , tSNE maps of different phenotypes of tumor-infiltrating lymphocytes after treatment, analyzed by FCM (n = 4 biologically independent samples). I , J , Quantitative analysis of matured dendritic cells and natural killer cells in draining lymph nodes by FCM (n = 4 biologically independent samples). K . Immunohistochemical images of PD-L1 in dissected tumors after treatment. L , Mean fluorescence intensity of PD-L1 in CD45-negative tumor cells from dissected tumors after treatment (n = 4 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Figure Legend Snippet: │ In vivo antitumor and degradation efficacy by PLT-TACs. A , Schematic schedule of the treatment and evaluation in BALB/c mice with 4T1 orthotopic tumor model. B , Tumor growth curves, C , images of dissected tumors, D , tumor weights, E , survival curves, F , body weight changes, G , tumor growth kinetics from the control and treatment groups (saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h ) at a dosage of 5 mg/kg (n = 6 biologically independent samples). H , tSNE maps of different phenotypes of tumor-infiltrating lymphocytes after treatment, analyzed by FCM (n = 4 biologically independent samples). I , J , Quantitative analysis of matured dendritic cells and natural killer cells in draining lymph nodes by FCM (n = 4 biologically independent samples). K . Immunohistochemical images of PD-L1 in dissected tumors after treatment. L , Mean fluorescence intensity of PD-L1 in CD45-negative tumor cells from dissected tumors after treatment (n = 4 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques Used: In Vivo, Control, Saline, Immunohistochemical staining, Fluorescence

│ Biodistribution and antitumor study by intravenous treatment. A , Schematic schedule of the intravenous treatment in BALB/c mice with 4T1 orthotopic tumor model. B , TEM image and DLS size distribution of LNP-TAC. C , The images of biodistribution at 24 h after treatment (n = 3 biologically independent samples). D , Hemodynamics analysis (n = 3 biologically independent samples). E , Tumor growth curves, F , images of dissected tumors, G , tumor weights, H , tumor growth kinetics after treatment (saline, LNP-TAC, and PLT-TAC m ) at a dosage of 5 mg/kg (n = 6 biologically independent samples). I , Immunohistochemical images of PD-L1 in tumors after treatment. J , Mean fluorescence intensity of PD-L1 in CD45-negative tumor cells from dissected tumors after the indicated treatment (n = 4 biologically independent samples). K , Hemolysis test (n = 3 biologically independent samples). L , Blood biochemical analysis (ALT, AST and CREA) after treatment (n = 4 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Figure Legend Snippet: │ Biodistribution and antitumor study by intravenous treatment. A , Schematic schedule of the intravenous treatment in BALB/c mice with 4T1 orthotopic tumor model. B , TEM image and DLS size distribution of LNP-TAC. C , The images of biodistribution at 24 h after treatment (n = 3 biologically independent samples). D , Hemodynamics analysis (n = 3 biologically independent samples). E , Tumor growth curves, F , images of dissected tumors, G , tumor weights, H , tumor growth kinetics after treatment (saline, LNP-TAC, and PLT-TAC m ) at a dosage of 5 mg/kg (n = 6 biologically independent samples). I , Immunohistochemical images of PD-L1 in tumors after treatment. J , Mean fluorescence intensity of PD-L1 in CD45-negative tumor cells from dissected tumors after the indicated treatment (n = 4 biologically independent samples). K , Hemolysis test (n = 3 biologically independent samples). L , Blood biochemical analysis (ALT, AST and CREA) after treatment (n = 4 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques Used: Saline, Immunohistochemical staining, Fluorescence

Cell Culture:

Article Title: High-resolution quantitative mapping of extracellular pH by ratiometric MRI with iron chelates in a tumor mouse model
Article Snippet: All cell culture media and solution were from Gibco, ThermoFisher Scientific, USA. .. The 4T1 breast tumor cell line (ATCC, USA, RRID: CVCL_0125) was cultured in RPMI-1640 medium supplemented with 10% heat-inactivated FBS and 1% penicillin/streptomycin (Gibco, ThermoFisher Scientific, USA). .. The BRL-3A hepatocytes (ATCC, USA, RRID: CVCL_0606) were cultured in DMEM medium supplemented with 10% heat-inactivated FBS and 1% penicillin/streptomycin.

Article Title: High-resolution quantitative mapping of extracellular pH by ratiometric MRI with iron chelates in a tumor mouse model.
Article Snippet: Cell culture and cell viability assay All cell culture media and solution were from Gibco, ThermoFisher Scientific, USA. .. The 4T1 breast tumor cell line (ATCC, USA, RRID: CVCL_0125) was cultured in RPMI-1640 medium supplemented with 10% heatinactivated FBS and 1% penicillin/streptomycin (Gibco, ThermoFisher Scientific, USA). .. The BRL-3A hepatocytes (ATCC, USA, RRID: CVCL_0606) were cultured in DMEM medium supplemented with 10% heat-inactivated FBS and 1% penicillin/streptomycin.

Modification:

Article Title: Chloroquine (CQ) exerts anti-breast cancer through modulating microenvironment and inducing apoptosis.
Article Snippet: a Department of Laboratory Medicine, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, China b Department of Surgical Oncology and Breast Surgery, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, China c Department of Medical Examination Center, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, China d Department of Immunology, College of Basic Medical Sciences, China Medical University, Shenyang, Liaoning 110122, China

Image Search Results

Journal: Bioactive Materials

Article Title: “Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation

doi: 10.1016/j.bioactmat.2026.01.011

Figure Lengend Snippet: │ Construction and characterization of PLT-TACs. A , Fast fabrication process of PLT-TACs via microfluidic device with a herringbone structure chip. B , TEM images and DLS size distribution of activated platelets (PLTs). C , TEM images and DLS size distribution of engineered activated platelets with medium density of anti-PD-L1 and P1 (PLT-TAC m ). D , SEM images of PLTs, E , SEM images of PLT-TAC m , white squares indicate zoom-in regions, red squares indicate morphology features on the platelet surface. F , Confocal images of Cy3-, Cy5-and Dio-labeled PLT-TACs. G , FCM analysis and 3D confocal visualization for verifying the co-installation of anti-PD-L1 and P1 in PLT-TACs. H , SDS-PAGE analysis of protein expression profiles in activated platelets and PLT-TAC m . I , FCM analysis of key platelet membrane proteins, CD41, CD47 and CD62P, expressed on PLT-TACs. J , AFM images of topography, height and Young's modulus mapping of PLTs and PLT-TAC m . Scale bars, 5 μm (n = 5 technical replicates). K , CLSM visualization of the localization of anti-PD-L1 and P1 in 4T1 cells after internalization for 3h, anti-PD-L1 was labeled with Cy5, P1 was labeled with Cy3, nuclei were stained with Hoechst 33342, and lysosomes were stained with lysotracker Green DND26. Scale bar, 10 μm. L , Fluorescence intensity profiles along the line drawn in K (n = 3 biological replicates).

Article Snippet: Breast tumor cell line 4T1 and melanoma tumor cell line B16F10 were purchased from the American Type Culture Collection (ATCC), and cultured in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin.

Techniques: Labeling, SDS Page, Expressing, Membrane, Staining, Fluorescence

Journal: Bioactive Materials

Article Title: “Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation

doi: 10.1016/j.bioactmat.2026.01.011

Figure Lengend Snippet: │ Surface binding, internalization, and PD-L1 degradation. A , Schematic illustration of the trafficking route of PLT-TACs. B , Fluorescence intensity analysis and histogram plots of PD-L1 binding to 4T1 cell surface after treatment with free anti-PD-L1, PLT-anti-PD-L1 or different PLT-TACs for 30min at 4 °C, measured by FCM (n = 3 biologically independent samples). C , Fluorescence intensity analysis and histogram plots of internalized PLT-anti-PD-L1 or different PLT-TACs in IFN-γ pre-treated 4T1 cells for 1, 2, or 4 h at 37 °C, measured by FCM (n = 3 biologically independent samples). D , Visualization of internalization of PLT-anti-PD-L1 or different PLT-TACs in IFN-γ pre-treated 4T1 cells after treatment for 2 h, observed by 40 × CLSM. Scale bar, 10 μm. E , Redistribution of PLT-TAC m membrane after treatment for 2 h or 4 h, observed by oil 63 × CLSM. Platelets were stained with Dio. Scale bar, 5 and 20 μm. F , Visualization and fluorescence intensity profiles along the line of colocalization of PLT-anti-PD-L1 or different PLT-TACs with lysosomes in IFN-γ pre-treated 4T1 cells after treatment for 4 h, observed by oil 63 × CLSM. Lysosomes were stained with lysotracker Green DND26. Scale bar, 10 μm. G , Mean fluorescence intensity of levels of PD-L1 in IFN-γ pre-treated 4T1, measured by FCM after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). H , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). I , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m with different concentration (0, 0.025, 0.25, 1.25, 2.5, 5 μM) for 36 h (n = 3 biologically independent samples). J , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) with different time (0, 6, 12, 24, 36, 48 h) (n = 3 biologically independent samples). K , Immunofluorescence of surface PD-L1 in IFN-γ pre-treated 4T1 cells treated with PLT-TAC m (5 μM) for 36 h. Scale bar, 20 μm. L , Visualization of PD-L1 degradation in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) for 0 h or 24 h. Scale bar, 5 μm. M , Western blot analysis of CD24, CD71 and CD47 in IFN-γ pre-treated 4T1 after treated with PLT-TAC m (5 μM) for 36 h (n = 3 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques: Binding Assay, Fluorescence, Membrane, Staining, Saline, Western Blot, Concentration Assay, Immunofluorescence

Journal: Bioactive Materials

Article Title: “Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation

doi: 10.1016/j.bioactmat.2026.01.011

Figure Lengend Snippet: │ Internalization mechanism analysis of PLT-TACs. A, Mutated P1 peptide (M1) designed using ProteinMPNN. B , Fluorescence intensity analysis of internalized M1-modified PLT-TAC m (M1) for 1, 2, or 4 h at 37 °C, measured by FCM (n = 3 biologically independent samples). C , Western blot analysis of PD-L1 in IFN-γ pre-treated 4T1 cells after treated with PLT-TAC m (M1) and PLT-TAC m (at an equivalent amount of 5 μM) for 36 h (n = 3 biologically independent samples). D , Dynamic internalization process of PLT-TAC m at different time points, observed by confocal microscopy. Scale bar = 5 μm. E , Visualization and fluorescence intensity profiles along the line of the colocalization of PLT-TACs with endo/lysosomes in IFN-γ pre-treated 4T1 cells after treated for 2 h or 4 h, observed by oil 63 × CLSM (Early endosomes were stained by EEA1, lysosomes were stained by LAMP1, anti-PD-L1 was labeled with Cy5, nuclei were stained with Hoechst 33342.). Scale bar, 10 μm (n = 3 biologically independent samples). F , Visualization of the amount of lysosome biogenesis in IFN-γ pre-treated 4T1 cells after treated with PLT-anti-PD-L1 or PLT-TACs for 2 h or 4 h, observed by oil 63 × CLSM. Lysosomes were stained by LAMP1. Scale bar, 10 μm. G , Quantification of LAMP1 positive puncta in saline and PLT-TAC m treated cells after 4 h treatment. H , Western blot analysis of LAMP1 after treated with saline and PLT-TAC m for 36 h. I , Visualization of the colocalization of PLT-anti-PD-L1 and PLT-TAC m with LC3-II-labeled autophagosomes (yellow box) in IFN-γ pre-treated 4T1 cells after treatment for 4 h, observed by oil 63 × CLSM. Autophagosomes were stained by LC3-Ⅱ, anti-PD-L1 was labeled with Cy5, nuclei were stained with Hoechst 33342. Scale bar, 10 μm. J , Western blot analysis of LC3-I and LC3-II in after treated with PLT-anti-PD-L1 and PLT-TAC m for 36 h. K , Visualization of localization of TFEB and statistical analysis of cells with nuclear localization of TFEB after treated with PLT-anti-PD-L1 and PLT-TAC m for 36 h. Scale bar, 20 μm. (n = 3 biologically independent samples). L , Fluorescence intensity of internalized PLT-anti-PD-L1 and PLT-TAC m in IFN-γ pre-treated 4T1 cells after treatment for 2 h in presence of chlorpromazine, nystatin, colchicine, and cytochalasin D, measured by FCM (n = 3 biologically independent samples). M , Visualization of internalization of PLT-TACs and lysosome biogenesis in IFN-γ pre-treated 4T1 cells after treatment for 2 h in presence of chlorpromazine, nystatin, colchicine, and cytochalasin D, observed by oil 63 × CLSM. Scale bar, 10 μm. N, SEM images of internalization of PLT-TAC m by IFN-γ pre-treated 4T1 cells. Scale bar, 10 μm. Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques: Fluorescence, Modification, Western Blot, Confocal Microscopy, Staining, Labeling, Saline

Journal: Bioactive Materials

Article Title: “Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation

doi: 10.1016/j.bioactmat.2026.01.011

Figure Lengend Snippet: │ Efficacy and versatility of PLT-TACs. A , Molecular mechanism underlying PD-L1 degradation-induced apoptosis. B , C , FCM analysis and quantification of cell apoptosis after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h, followed by annexin V-FITC/PI staining (n = 3 biologically independent samples). D , E , Images and statistical analysis of wound healing assay in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h. Scale bar, 200 μm (n = 3 biologically independent samples). F , G , Images and statistical analysis of transwell cell invasion assay in IFN-γ pre-treated 4T1 cells after treated with saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h (at an equivalent amount of 5 μM) for 24 h. Scale bar, 200 μm (n = 3 biologically independent samples). H , Structures of CD71 and T7 peptide and the binding site of CD71/T7 complex, calculated by AlphaFold 3. I , Construction and characterization of PLT-TAC T7 by TEM and confocal microscopy. J , Mean fluorescence intensity of levels of surface CD71 in 4T1 cells after treated with saline, PLT-anti-PD-L1, or different PLT-TAC T7 (at an equivalent amount of 10 μM) for 36 h, measured by FCM (n = 3 biologically independent samples). K , Western blot analysis of CD71 in IFN-γ pre-treated 4T1 cells after treated with saline, and different PLT-TAC T7 (at an equivalent amount of 10 μM) for 36 h (n = 3 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques: Saline, Staining, Wound Healing Assay, Invasion Assay, Binding Assay, Confocal Microscopy, Fluorescence, Western Blot

Journal: Bioactive Materials

Article Title: “Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation

doi: 10.1016/j.bioactmat.2026.01.011

Figure Lengend Snippet: │ In vivo antitumor and degradation efficacy by PLT-TACs. A , Schematic schedule of the treatment and evaluation in BALB/c mice with 4T1 orthotopic tumor model. B , Tumor growth curves, C , images of dissected tumors, D , tumor weights, E , survival curves, F , body weight changes, G , tumor growth kinetics from the control and treatment groups (saline, PLT-anti-PD-L1, PLT-TAC l , PLT-TAC m or PLT-TAC h ) at a dosage of 5 mg/kg (n = 6 biologically independent samples). H , tSNE maps of different phenotypes of tumor-infiltrating lymphocytes after treatment, analyzed by FCM (n = 4 biologically independent samples). I , J , Quantitative analysis of matured dendritic cells and natural killer cells in draining lymph nodes by FCM (n = 4 biologically independent samples). K . Immunohistochemical images of PD-L1 in dissected tumors after treatment. L , Mean fluorescence intensity of PD-L1 in CD45-negative tumor cells from dissected tumors after treatment (n = 4 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques: In Vivo, Control, Saline, Immunohistochemical staining, Fluorescence

Journal: Bioactive Materials

Article Title: “Artificial platelet injection system”: a plug-and-play platelet-based lysosome-targeting chimera for targeted protein degradation

doi: 10.1016/j.bioactmat.2026.01.011

Figure Lengend Snippet: │ Biodistribution and antitumor study by intravenous treatment. A , Schematic schedule of the intravenous treatment in BALB/c mice with 4T1 orthotopic tumor model. B , TEM image and DLS size distribution of LNP-TAC. C , The images of biodistribution at 24 h after treatment (n = 3 biologically independent samples). D , Hemodynamics analysis (n = 3 biologically independent samples). E , Tumor growth curves, F , images of dissected tumors, G , tumor weights, H , tumor growth kinetics after treatment (saline, LNP-TAC, and PLT-TAC m ) at a dosage of 5 mg/kg (n = 6 biologically independent samples). I , Immunohistochemical images of PD-L1 in tumors after treatment. J , Mean fluorescence intensity of PD-L1 in CD45-negative tumor cells from dissected tumors after the indicated treatment (n = 4 biologically independent samples). K , Hemolysis test (n = 3 biologically independent samples). L , Blood biochemical analysis (ALT, AST and CREA) after treatment (n = 4 biologically independent samples). Data are presented as mean ± SD. p values were determined by one-way ANOVA test. ns, no significance; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Techniques: Saline, Immunohistochemical staining, Fluorescence

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breast tumor cell line 4t1 (ATCC)

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