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. 2019 Nov 22;56(1):258–272. doi: 10.3892/ijo.2019.4918

Figure 1.

Figure 1

rhIL-6 recapitulates many of the effects of CAF-CM on protein expression in MCF-7 cells. (A) Antibody arrays of CAF-CM and control medium. (B) Effects of CAF-CM and rhIL-6 on relative RNA expression on Bcl-3, IGFBP5 and STAT3 as measured by RT-qPCR. (C) Western blot analyses of protein extracts derived from cells treated with either CAF-CM or rhIL-6 (0.3 µg/ml) or of untreated cells (control). Cells were incubated for 3 days before plasma membrane (PM), cytosolic (CE) and nuclear (NE) proteins were extracted. (D) Effect of a Bcl-3-specific siRNA (siBcl3) on the protein expression of Bcl-3, c-Myc and p50 (NF-κB) as determined by western blot analyses of nuclear protein extracts. (E) Effects of CAF-CM and rhIL-6 (0.3 µg/ml) on the relative RNA expression of ALDH1A1 and ALDH1A3 as measured by RT-qPCR. (F) Effect of CAF-CM and rhIL-6 (0.1 and 0.3 µg/ml) on the relative RNA expression of 11 CAF-CM-responsive genes. (B, E and F) Each bar represents the average values of 3 independent experiments. Statistical analyses were carried out by ANOVA and the Bonferroni post-hoc test. Asterisk(s) indicate statistical significance in comparison to the control. *P<0.05, **P<0.01, ***P<0.005 and ****P<0.001. rhIL-6, recombinant human interleukin 6; CAF-CM, carcinoma-associated fibroblast-conditioned medium; ALDH1A1, aldehyde dehydrogenase 1 family, member A1; ALDH1A3, aldehyde dehydrogenase 3 family, member A1; STAT3, signal transducer and activator of transcription 3.