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. Author manuscript; available in PMC: 2016 Mar 9.
Published in final edited form as: Dev Cell. 2015 Mar 9;32(5):640–651. doi: 10.1016/j.devcel.2015.01.011

Figure 1. Impact of Cell Density on TGF-β Signaling.

Figure 1

HaCaT keratinocytes, 1205Lu melanoma cells, and EpH4 mouse mammary epithelial cells were grown in either low (LD) or high (HD) density conditions prior to TGF-β (5 ng/ml) stimulation.

(A) Quantitative RT-PCR analysis of PAI-1 expression after a 24-hr TGF-β treatment. Results are expressed as -fold induction by TGF-β in each culture condition and are the mean ± SD from three independent experiments, each measured in triplicate.

(B) Effect of TGF-β on SMAD3/4-specific transcription. Results are expressed as -fold activation of transiently transfected (CAGA)9-MLP-luc activity 18 hr after TGF-β addition to the cultures. Results are the mean ± SD of two independent experiments, each performed with triplicate samples.

(C) Western analysis of P-SMAD3 levels without or with 30 min TGF-β stimulation. Actin levels were measured as a control for the specificity of P-SMAD3 changes under each experimental condition. Results from one representative of several independent experiments are shown.