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. 2015 Mar 12;6(13):11009–11022. doi: 10.18632/oncotarget.3557

Figure 4. TGFβ1 enhanced the smad2 signaling pathway in mLRG-overexpressing LLC.

Figure 4

a. Western blot analysis shows the phosphorylation of smad2 and smad1/5/8 in LLC/mLRG-3 and LLC/CV-8 cells treated with TGFβ1. After 6 h of serum starvation, cells were treated with or without TGFβ1 (1.0 ng/mL) for 10 or 30 min. b. Western blot analysis shows the phosphorylation of smad2 in LLC/mLRG-3 and LLC/CV-8 cells treated with or without SB431542 (10 μM) and TGFβ1. Cells were treated with SB431542 (10 μM) or DMSO (vehicle) for 3 h and with TGFβ1 (1.0 ng/mL) for 30 min. c. Quantitative real-time PCR analysis shows PAI-1 gene and Id-1 gene expression with or without stimulation with TGFβ1 (1.0 ng/mL) for 3 h in LLC/mLRG-3 and LLC/CV-8 cells after 6 h of serum starvation. Quantitative real-time PCR threshold values for the target genes were normalized against the level of HPRT1. d. Quantitative real-time PCR analysis shows TIEG gene expression with or without stimulation with TGFβ1 (1.0 ng/mL) for 3 h in LLC/mLRG-3 and LLC/CV-8 cells after 6 h of serum starvation. Quantitative real-time PCR threshold values for the target genes were normalized against the level of HPRT1. e. Western blot analysis shows Bcl-2 and Bcl-xL protein in LLC/mLRG-3 and LLC/CV-8 cells treated with TGFβ1. Cells were cultured in 6-well plates with or without stimulation with TGFβ1 (1.0 ng/mL). After culture for 24 h, cell lysates were collected. Each value is the average ± standard deviation. *P < 0.01.