Figure 6

Co-cultures of ATC cells with fibroblasts induce the protein expression of CAF-markers in human thyroid fibroblasts. Effects of CM from 8505c cells on CAFs markers in human thyroid fibroblasts (A–D). (A) Immunoblot analysis of PDGFR-β (a) and α-SMA (b) in human thyroid fibroblasts, grown under normal conditions (lanes 1 and 2) or exposed to CM derived from 8505c cells for 48 h (lanes 3 and 4). The figure shows a representative Western Blot of 2 independent experiments (n = 2) with duplicate samples for each experimental group. (B) Quantification of relative expression of PDGFR-β and α-SMA proteins in human thyroid fibroblasts after using GAPDH as loading control. [Full blots are shown in the Supplemental Information, Full Original Blots-VI]. Data are expressed as mean ± SD. (C) Immunoblot analysis in human thyroid fibroblast after 48 h of co-cultures with ATC cells. Immunoblot analysis of α-SMA (a) in mono-cultures of human thyroid fibroblast (lanes 1 and 2) or after co-cultures with 8505c cells (lanes 3 and 4) in 12-well transwell system, for 48 h. The figure shows a representative Western Blot of 2 independent experiments (n = 2) with duplicate samples for each experimental group. (D) Quantification of relative expression of α-SMA protein in human thyroid fibroblasts after using GAPDH as loading control (Full blots are shown in the Supplemental Information, Full Original Blots-VI). Data are expressed as mean ± SD. (E) Immunoblot analysis of vimentin in human thyroid fibroblasts grown under normal conditions (lanes 1 and 4) or exposed to CM derived from 8505c for 48 h (lanes 2 and 3) and 72 h (lanes 5 and 6). The figure shows a representative Western Blot of 2 independent experiments (n = 2) with duplicate samples for each experimental group. (F) Quantification of relative expression of vimentin protein in human thyroid fibroblasts after using GAPDH as loading control. The same blot was stripped and re-blotted using anti-GAPDH antibody for loading control (Full blots are shown in the Supplemental Information, Full Original Blots-VI). Data are expressed as mean ± SD. (G) Immunoblot analysis of GLUT-1 in human thyroid fibroblasts grown under normal conditions (lanes 1 and 4) or exposed to CM derived from 8505c for 48 h (lanes 2 and 3) and 72 h (lanes 5 and 6). The figure shows a representative Western Blot of 2 independent experiments (n = 2) with duplicate samples for each experimental group. (H) Quantification of relative expression of GLUT-1 protein in human thyroid fibroblasts after using GAPDH as loading control. The same blot was stripped and re-blotted using anti-GAPDH antibody for loading control. [Full blots are shown in the Supplemental Information, Full Original Blots-VI]. Data are expressed as mean ± SD.