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. Author manuscript; available in PMC: 2010 Jun 8.
Published in final edited form as: Biochem J. 2009 Apr 15;419(2):485–495. doi: 10.1042/BJ20081434

Figure 2. Changes in KLF6 alternative splicing after TGF-β1 treatment in different cells.

Figure 2

(A and B) HEK-293T cells were co-transfected with pCD105 (− 50/+ 350)-pXP2 (pEndoglin, A) or pGL-Col1 (pCollagen, B) and 1 μg of either WT KLF6 (KLF6wt) (black bar 2), KLF6 Sv1 (Sv1) (black bar 3), KLF6 Sv2 (Sv2) (black bar 4) or a mixture of 0.5 μg of WT and 0.5 μg of Sv1 (black bar 5)/Sv2 (black bar 6). For the first two bars in (A), endogenous KLF6 was knocked down with KLF6 siRNA (white bar). In addition, the effect of TGF-β treatment (10 ng/ml) was assessed in hatched bars 1–4 in (A). The results are given in arbitrary units of LUC activity. Representative results obtained from three different experiments with replicable results are shown. *Statistical significance at least P < 0.05 between control (pEndoglin, pCollagen) and KLF6 co-transfected. In case of the white bar (siRNA), the asterisk (*) means that the value was statistically significant (P<0.05) compared with the corresponding black bar 2. (C–E) After THP-1 (C), HUVEC (D) and HepG2 (E), cells were treated with 10 ng/ml TGF-β1 for the various times indicated and cell lysates were prepared. Total RNA was extracted from each cell lysate, and levels of WT KLF6 and its Svs in each sample were measured using real-time PCR. To calculate the fold change in mRNA levels of KLF6 alternative splicing, the fold change in mRNA levels of total KLF6 (WT KLF6 plus alternatively spliced KLF6 transcripts) was divided by the fold change in wild-type KLF6 alone. *Statistical significance at least P < 0.05.