Protocol detail

Exosome Uptake and Stat3 Activation in OSCC

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Exosome uptake by macrophages was examined as previously reported [8 (link)]. Differently treated OSCC cells were fixed, permeabilized and blocked as previously reported [15 (link)]. Then, the cells were incubated with anti-phospho-Stat3 (Y705) antibody (Cell Signaling Technology, USA) overnight at 4 °C followed by secondary antibody (Life, USA). Cellular nuclei were stained with DAPI (Roche, USA). Fluorescently labelled OSCC cells were examined using a ZEISS fluorescent imaging microscope (ZEISS, German).

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You Y., Tian Z., Du Z., Wu K., Xu G., Dai M., Wang Y, & Xiao M. (2022). M1-like tumor-associated macrophages cascade a mesenchymal/stem-like phenotype of oral squamous cell carcinoma via the IL6/Stat3/THBS1 feedback loop. Journal of Experimental & Clinical Cancer Research : CR, 41, 10.

Publication 2022

DAPI fluorescent imaging microscope

Antibody Cells Cellular nuclei Dapi Exosome Macrophages Microscope Stat3

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Variable analysis

independent variables

Differently treated OSCC cells

dependent variables

Exosome uptake by macrophages
Phospho-Stat3 (Y705) expression in OSCC cells

control variables

Fixation, permeabilization, and blocking of OSCC cells as previously reported [15]
Incubation of OSCC cells with anti-phospho-Stat3 (Y705) antibody overnight at 4 °C
Incubation of OSCC cells with secondary antibody
Staining of cellular nuclei with DAPI

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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