Gremlin is significantly overexpressed in lung AD but not lung SCC.

<p>A Taqman qRT-PCR was performed using GREM1 probe and a GAPDH control. Data are represented as GREM1 expression normalized to GAPDH expression for each sample. Gremlin expression was higher in AD specimens than matched normals (N = 96, p = 5.438×10⁻²²). There was no significant change in expression in the SCC samples (N = 65, p = 0.705).</p

mRNA expression profile of <i>FBXW7α</i> AS forms in human normal tissues.

<p>(A) <i>FBXW7α</i> AS forms expression levels were detected by semi-quantitative RT-PCR using different pairs of primers. (B) The levels of Spli1&2 and Spli3-5 expression were quantified from the experiment shown in (A) lower panel. Normal tissues include: 1-esophagus, 2-adipose, 3-heart, 4-bladder, 5-kidney, 6-brain, 7-liver, 8-lung, 9-cervix, 10-colon, 11-spleen, 12-testes, 13-thymus, 14-thyraoid, 15-trachea, 16-small intestine, 17-skeletal muscle, 18-prostate, 19-placental, 20-ovary, 21-breast. “M” represents DNAs ladder Marker.</p

5′-UTRs of <i>FBXW7α</i> determine the translational efficiency.

<p>(A) Effect of <i>FBXW7α</i> 5′-UTR variants on LUC activity. LUC reporter constructs, which contained each <i>FBXW7α</i> 5′-UTR upstream of the LUC reporter gene in the pGL3 vector, were transfected into Hela cells. The LUC activity was measured as the Firefly/Renilla ratio. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049453#s2" target="_blank">Results</a> represent the data from three independent experiments. Each experiment was performed in triplicate. Mean data and standard deviations are shown. *p<0.05 in comparison to LUC-ctr. (B) LUC transcriptional levels were examined by qRT-PCR. The LUC mRNA contents were normalized to the Renilla mRNA contents for all samples and the relative LUC mRNA for pGL3p (empty vector, Promega) was arbitrarily considered to be 1 (control). (C) Effect of 5′-UTRs on the FBXW7α protein levels. Immunoblot was developed with anti-HA antibody from extracts with indicated construct. The intensity of pcDNA3.1(+) containing FBXW7α gene (control) was considered as 1. GFP construct was used as transfection efficiency control, and β–actin as loading control. All transfections were performed in triplicate, and the bars indicate the standard deviation. *p<0.05 in comparison to control. (D) cDNA frangments for FBXW7 (upper panel), GFP (middle panel) and GAPDH (lower panel) were specifically amplified by RT-PCR from Hela cells transfected with indicated constructs.</p

Mutational analysis of <i>FBXW7</i> in primary prostate, bladder, and kidney cancers from the Chinese patients.

<p>(A) RT-PCR analysis of <i>FBXW7</i>, RT–PCR products containing deletions are indicated by asterisk. (B–D) sequencing confirmed the deletion in two bladder cancers (B and C), and a kidney cancer (D). (E) Representative sequence traces showing the mutations of <i>FBXW7</i>.</p