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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of <t>CTCs</t> with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.
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(A) Representative immunofluorescence images of CTCs with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.

Journal: The Journal of Liquid Biopsy

Article Title: Novel applications of liquid Biopsy: Comprehensive methodology for circulating biomarker exploration in peripheral blood

doi: 10.1016/j.jlb.2025.100307

Figure Lengend Snippet: (A) Representative immunofluorescence images of CTCs with different extraction methods. Co-localization of STING (green, Alexa Fluor 488) and vimentin (red, Alexa Fluor 647). Nuclei were stained with DAPI (blue). Magnification 63X. (B) Representative SEM images of the isolated exosomes (scale bar = 2 μm). (C) Western blot of exosomal markers CD81 and CD63. The isolated exosomes with different protein concentrations (EXO#1 = 40 μg and EXO#2 = 20 μg) were successfully obtained from the culture supernatants of PBMCs from SCLC patients. Red line indicated the band of each protein on the gel. Original western blots are presented in File S1. (D) Stacked bar plots showing the log10-transformed positive control read counts across the samples. (E) Violin plot displaying the distribution of the log of the un-normalized gene counts for all the samples.

Article Snippet: MACS® Tumor Cell Isolation Kit depletes tumor-associated cells without harming CTCs; we modified the protocol by omitting Non-Tumor Cell Depletion Cocktail B to preserve mesenchymal CTCs (Order no. 130-108-339, www.miltenyibiotec.com ).

Techniques: Immunofluorescence, Extraction, Staining, Isolation, Western Blot, Transformation Assay, Positive Control