Figure 2.

Exosomes Secreted from ESCC cells Induce NF Activation into CAFs

(A) Representative morphology of NFs and CAFs isolated from patients with ESCC. (B) Representative immunohistochemical images of the NF marker (vimentin) and CAF marker (α-SMA) expression in ESCC tissues and adjacent normal tissues. Scale bars, 50 μm. (C) Western blot analysis of the expression of vimentin, α-SMA, and FAP in NFs and CAFs isolated from two pairs of esophageal cancer tissues and normal esophageal tissues. (D) Immunofluorescence staining of vimentin, α-SMA, and FAP in NFs and CAFs. Scale bars, 50 μm. (E) NFs were incubated with PKH26-labeled exosomes from KYSE450, TE12, or Het-1a cells for 24 h, and the red exosome signal was detected by fluorescence microscopy. Scale bars, 50 μm. (F) NFs were treated with KYSE450, TE12, or Het-1a cell-secreted exosomes (25 μg/mL) for 48 h. Fluorescence microscopy was used to detect the expression of α-SMA (green) and FAP (red) in NFs. Scale bars, 50 μm. (G) Western blot analysis of α-SMA and FAP expression in NFs treated with exosomes secreted from ESCC cells (25 μg/mL).