Fig. 2.

T-KSR2 is expressed exclusively in testes and mature sperm. (A) Primer pairs used for detecting KSR2 mRNA. F9 and 207R selectively detect full-length KSR2. F5UTR and R490 primer pairs selectively detect T- KSR2. F610 and Rstop primer pairs detect both mRNAs. T-KSR2 exon1t which can be recognized only by F5UTR primer is in gray. (B) RT-PCR was performed with primers described in panel A. RNA was isolated from the indicated tissues from 8 to 12 week old wild type C57BL/6 mice. BAT: brown adipose tissue, WAT: white adipose tissue, MUS: Quadriceps muscle. (C) Male reproductive tracts were dissected from 12 week old mice. RNA was purified and RT-PCR was done with the primer pairs illustrated in panel A. (D) RT-PCR was performed with primers detecting somatic cell marker E-cadherin and round germ cell marker c-kit. NRT: no reverse transcriptase control for the previous sample. (E) Blood samples were tested for T-KSR2 expression. (F) RT-PCR was performed with purified sperm (p-Sperm) treated with hypotonic buffer for detecting T-KSR2, protamine-1, and E-cadherin expression. Yeast tRNA was used as a negative control. NRT: no reverse transcriptase control for testes.