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Figure 4 | Molecular Cancer

Figure 4

From: Long-term reduction of T-cell intracellular antigens leads to increased beta-actin expression

Figure 4

The ablation of TIA1 gene in murine embryonic fibroblasts increases the expression of mouse β-actin protein without affecting the levels of mRNA. (A) A chimeric GFP RNA containing β-actin 3′-UTR reproduces the regulation of the endogenous human β-actin mRNA in control (c), TIA1, TIAR and HuR-knocked down HeLa cells. The relative expression levels of GFP protein versus endogenous α-tubulin levels as well as the levels of reporter GFP mRNAs versus endogenous GAPDH mRNA were quantified by western blot analysis and QPCR, respectively. The quantifications are represented by histograms where white and black bars correspond to the GFP protein/RNA expression from GFP and GFP-β-actin 3′-UTR transfected plasmids, respectively. (B) Effect of the disruption of mouse TIA1 gene on β-actin expression in murine embryonic fibroblasts (MEF). Protein extracts and cytoplasmic RNAs from normal murine embryonic fibroblasts (WT MEFs) or knocked out for TIA1 (TIA1 KO MEFs) were purified and processed for western blot analysis with anti-β-actin, anti-TIA1, anti-TIAR, anti-HuR, anti-U2AF65 and anti-α-tubulin antibodies, or for QPCR analysis with mouse β-actin and GAPDH specific-mRNA primers, respectively. Molecular weight markers for protein (kDa) are indicated on the left. The identities of protein bands are indicated on the right by arrowheads. The represented values by histograms were normalized and are expressed relative to control (c), whose value is fixed arbitrarily to 1.

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