Figure 1. Detecting the expression of FoxO6 at both mRNA and protein levels in various tissues.
(A) The expression of FoxO6 mRNA in various mouse tissues (N=3) was determined by qRT-PCR. The expression level of FoxO6 in liver was arbitrarily set as 1-fold and compared by the levels in other tissues.**, P<0.01 as compared with that of liver. (B) In vitro translated 35S-FoxO6 was detected with either auto-radiography (lower panel) or Western blot (upper panel) using home-made polyclonal antibody. FoxO6-MCS is a chimaeric protein containing FoxO6 CDS and the multiple cloning sites (from BamHI to XbaI) of pCDNA3 vector. Lysate: reticulocyte total lysate used for translation. *, non-specific band caused by haemoglobin in the reticulocyte. (C) GFP fused FoxOs were expressed in HEK-293 cells and their expression were detected by GFP antibody (lower panel) or FoxO6 antibody (upper panel). Total lysate (50 μg) from mouse and primate cells (D) or various mouse tissues (F and G) were examined for their expression of FoxO6 by Western blot. The signals of Gapdh serve as input control. (E) Alignment of FoxO6 cDNA sequences (1603–1622) reported by IMAGE Consortium, Smidt's group, and our group (C2C12). The corresponding genomic sequence is also aligned. Numbering is according to Smidt's clone. GC, gastrocnemius; EDL, extensor digitorum longus; TA, tibialis anterior; Quad, quadriceps.