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. 2019 May 29;9:8028. doi: 10.1038/s41598-019-44361-6

Figure 2.

Figure 2

ATC cells-derived CM induced the protein expression of CAF-markers in human fibroblasts. (A) Schematic representation of CM preparation from ATC cells and treatment of MRC-5 cells. (B–F) Effects of CM from ATC cells on CAFs markers in MRC-5 cells. Immunoblot analysis of PDGFR-β in MRC-5 cells grown under normal conditions (control) or exposed to CM derived from ATC cells, 8505c and KTC-2, for 24 h and 48 h (B). The figure shows a representative Western Blot of 3 independent experiments (n = 3) with duplicate samples for each experimental group. Quantification of relative expression of PDGFR-β protein in human fibroblasts after using GAPDH as loading control. The blot was cut and blotted using anti-GAPDH antibody. [(C) full blot is shown in the Supplemental Information, Full Original Blots-II]. Data are expressed as mean ± SD. (D) Concentrations of platelet derived growth factors in the CM of normal thyroid, MRC-5, 8505c and KTC-2 cells determined by ELISA as described in Methods and Materials. Data are expressed as mean ± SD of 2 independent experiments (n = 2) with triplicate samples for each experimental group; *Not detectable. (E) Immunoblot analysis of α-SMA in MRC-5 cells grown under normal conditions (control) or exposed to CM derived from ATC cells, 8505c and KTC-2, for 24 h and 48 h. The figure shows a representative Western Blot of 3 independent experiments (n = 3) with duplicate samples for each experimental group. (F) Quantification of relative expression of α-SMA protein in human fibroblasts after using GAPDH as loading control. The blot was stripped and re-blotted using anti-GAPDH antibody (full blot is shown in the Supplemental Information, Full Original Blots-II). Data are expressed as mean ± SD. *p < 0.05 and **p < 0.005. ns, not significant.