Figure 4
LXN is directly upregulated by the differentiation agent atRA in human prostate epithelial cells. (A) Significant induction of LXN (30.57 ± 3.011-fold p = 0.0006) and HOXA1 (3.65 ± 0.3421-fold p = 0.0015) mRNA expression in PNT1A cells 8 hrs after treatment with 1 µM atRA. RT-PCR analysis from 3 independent experiments. (B) A time-course showing significant induction of LXN protein expression from 24 hrs after treatment with 1 µM atRA. See Supplementary Fig. S10A,B for uncropped blots. (C) Protein half-life assay to determine the stability of LXN over a 24 hr time-course following treatment of PNT1A cells with 10 µM of the protein synthesis inhibitor anisomycin. The relatively unstable protein p53 serves as a positive control. See Supplementary Fig. S10C,D for uncropped blot images. (D) Dose dependent induction of LXN protein expression following treatment with 500 nM or 1 µM atRA. See Supplementary Fig. S10D,E for uncropped blot images. (E) Induction of LXN and HOXA1 mRNA expression over an 8 hr time-course following treatment with 1 µM atRA in PNT1A cells (n = 3). (F) Significant induction of HOXA1 (4.83-fold ± 1.26-fold p = 0.0158) and LXN (3.73 ± 0.57-fold p = 0.0015) mRNA expression in primary human prostate epithelial cells 24 hrs after treatment with 500 nM atRA (n = 9). (G) Assessment of LXN protein expression 72 hrs after treatment with 500 nM atRA in 9 individual primary prostate epithelial cell samples. Data obtained from 3 separate blots (indicated by hatched lines) (See Supplementary Fig. S11 for full sized uncropped blots and multiple exposures). Scanning densitometry of (F) revealed LXN to be significantly induced 2.54 ± 0.34- fold p = 0.0313 (n = 9).
