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Core@shell nanocarriers undergo intracellular endosomal trafficking: a–c) Representative confocal microscopy images of HCT116 cells either untreated (0 h) or treated with 60.53 μg mL −1 DUT647‐labeled REF‐NC (margenta) for 6 or 24 h. Cells were immunostained for the endosomal markers <t>EEA1</t> (a, green, early endosomes), RAB5 (b, green, early endosomes), and LAMP1 (c, green, lysosomes). d) Quantification of DUT647‐labeled REF‐NC colocalization with endosomal markers at different time points, expressed as the percentage of REF‐NC signal colocalizing with each marker. Data represent three independent experiments. e) Schematic representation of intracellular endosomal compartments, indicating key markers: early endosome (EEA1, RAB5), and lysosome (LAMP1).
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Core@shell nanocarriers undergo intracellular endosomal trafficking: a–c) Representative confocal microscopy images of HCT116 cells either untreated (0 h) or treated with 60.53 μg mL −1 DUT647‐labeled REF‐NC (margenta) for 6 or 24 h. Cells were immunostained for the endosomal markers <t>EEA1</t> (a, green, early endosomes), RAB5 (b, green, early endosomes), and LAMP1 (c, green, lysosomes). d) Quantification of DUT647‐labeled REF‐NC colocalization with endosomal markers at different time points, expressed as the percentage of REF‐NC signal colocalizing with each marker. Data represent three independent experiments. e) Schematic representation of intracellular endosomal compartments, indicating key markers: early endosome (EEA1, RAB5), and lysosome (LAMP1).
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Core@shell nanocarriers undergo intracellular endosomal trafficking: a–c) Representative confocal microscopy images of HCT116 cells either untreated (0 h) or treated with 60.53 μg mL −1 DUT647‐labeled REF‐NC (margenta) for 6 or 24 h. Cells were immunostained for the endosomal markers <t>EEA1</t> (a, green, early endosomes), RAB5 (b, green, early endosomes), and LAMP1 (c, green, lysosomes). d) Quantification of DUT647‐labeled REF‐NC colocalization with endosomal markers at different time points, expressed as the percentage of REF‐NC signal colocalizing with each marker. Data represent three independent experiments. e) Schematic representation of intracellular endosomal compartments, indicating key markers: early endosome (EEA1, RAB5), and lysosome (LAMP1).
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Core@shell nanocarriers undergo intracellular endosomal trafficking: a–c) Representative confocal microscopy images of HCT116 cells either untreated (0 h) or treated with 60.53 μg mL −1 DUT647‐labeled REF‐NC (margenta) for 6 or 24 h. Cells were immunostained for the endosomal markers <t>EEA1</t> (a, green, early endosomes), RAB5 (b, green, early endosomes), and LAMP1 (c, green, lysosomes). d) Quantification of DUT647‐labeled REF‐NC colocalization with endosomal markers at different time points, expressed as the percentage of REF‐NC signal colocalizing with each marker. Data represent three independent experiments. e) Schematic representation of intracellular endosomal compartments, indicating key markers: early endosome (EEA1, RAB5), and lysosome (LAMP1).
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Core@shell nanocarriers undergo intracellular endosomal trafficking: a–c) Representative confocal microscopy images of HCT116 cells either untreated (0 h) or treated with 60.53 μg mL −1 DUT647‐labeled REF‐NC (margenta) for 6 or 24 h. Cells were immunostained for the endosomal markers EEA1 (a, green, early endosomes), RAB5 (b, green, early endosomes), and LAMP1 (c, green, lysosomes). d) Quantification of DUT647‐labeled REF‐NC colocalization with endosomal markers at different time points, expressed as the percentage of REF‐NC signal colocalizing with each marker. Data represent three independent experiments. e) Schematic representation of intracellular endosomal compartments, indicating key markers: early endosome (EEA1, RAB5), and lysosome (LAMP1).

Journal: Small Science

Article Title: Macropinocytic Uptake and pH‐Responsive Endolysosomal Processing Drive Sustained Chemotherapeutic Efficacy of High‐Load Core@Shell Nanocarriers in Colorectal Cancer

doi: 10.1002/smsc.202500470

Figure Lengend Snippet: Core@shell nanocarriers undergo intracellular endosomal trafficking: a–c) Representative confocal microscopy images of HCT116 cells either untreated (0 h) or treated with 60.53 μg mL −1 DUT647‐labeled REF‐NC (margenta) for 6 or 24 h. Cells were immunostained for the endosomal markers EEA1 (a, green, early endosomes), RAB5 (b, green, early endosomes), and LAMP1 (c, green, lysosomes). d) Quantification of DUT647‐labeled REF‐NC colocalization with endosomal markers at different time points, expressed as the percentage of REF‐NC signal colocalizing with each marker. Data represent three independent experiments. e) Schematic representation of intracellular endosomal compartments, indicating key markers: early endosome (EEA1, RAB5), and lysosome (LAMP1).

Article Snippet: The following primary antibodies were used: polyclonal rabbit anti‐Rab5 (1:100, PA5‐29022, Invitrogen), Coralite Plus 488‐conjugated EEA1 monoclonal antibody (1:100, CL488‐68065, Proteintech), mouse anti‐LAMP1 monoclonal antibody (H4A3, Alexa Fluor 488‐conjugated, 1:100, MA5‐18121, Invitrogen), and rabbit anti‐ANKFY1 (1:100, HPA065849, Sigma‐Aldrich).

Techniques: Confocal Microscopy, Labeling, Marker