Figure 3

ATC cells-derived CM increased proliferation of MRC-5 cells. Proliferation of MRC-5 cells, incubated with 8505c-derived CM (A) or KTC-2-derived CM (B) for 24 h or 48 h, estimated by cell counting. Data are expressed as mean ± SD of 4 independent experiments (n = 4) with triplicate samples for each experimental group. Immunoblot analysis of p21 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 (C). 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 p21 protein in human fibroblasts after using GAPDH as loading control [(D) full blot is shown in the Supplemental Information, Full Original Blots-III]. The same blot was stripped and re-blotted using anti-GAPDH antibody for loading control. Immunoblot analysis of p27 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 (E). 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 p27 protein in human fibroblasts after using GAPDH as loading control [(F); full blot is shown in the Supplemental Information, Full Original Blots-III]. The same blot was cut and blotted using anti-GAPDH antibody for loading control. Immunoblot analysis of Src in MRC-5 cells grown under normal conditions (control) or exposed to CM derived from ATC cells, 8505c and KTC-2, for 24 h (G). The figure shows a representative Western Blot of 4 independent experiments (n = 4) with duplicate samples for each experimental group. Quantification of relative expression of pSrc/Total Src proteins in human fibroblasts [(H); full blot is shown in the Supplemental Information, Full Original Blots-III]. The same blot was stripped and re-blotted using anti-Total Src antibody. Data are expressed as mean ± SD. Immunoblot analysis of Akt 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 (I). 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 pAkt/Total Akt proteins in human fibroblasts [(J); full blot is shown in the Supplemental Information, Full Original Blots-III]. The same blot was stripped and re-blotted using anti-Total Akt antibody. Data are expressed as mean ± SD. *p < 0.05 and **p < 0.005. ns, not significant.