FIG. 4.
Expression of Sp110 and Sp110b mRNAs in human tissues and cell lines. (A) Northern blot analysis detecting Sp110 and Sp110b mRNAs in human tissues. (Upper panel) Ten-microgram portions of total RNAs from human brain, heart, liver, lung, kidney, bone marrow, placenta, thymus, testis, and spleen were analyzed by using a 32P-labeled Sp110b fragment as a probe. Positions of molecular size markers are indicated on the left. (Lower panel). Ethidium bromide staining of 28S rRNA served as a loading control. (B) Semiquantitative RT-PCR analysis of Sp110 and Sp110b mRNAs in human tissues and cell lines. (Upper panel) cDNA fragments of Sp110 and Sp110b mRNAs were simultaneously amplified by RT-PCR using a common sense primer and two antisense primers specific to Sp110 and Sp110b mRNAs, respectively. To evaluate the quantifying ability of this system, mixtures of in vitro-synthesized Sp110 and Sp110b RNAs at various concentrations (given above the gel, in femtograms) were used as templates (lanes 1 to 9). Total RNAs from human kidneys (lane 10) and spleens (lane 11) and from MCF-7 (lane 12), Huh-7 (lane 13), Jurkat (lane 14), and HeLa (lane 15) cells were examined. (Lower panel) As an internal control, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was detected by a similar protocol (lanes 10 to 15). RTase(−), experimental control treated like other samples, but without reverse transcriptase.