Human normal testis total RNA was obtained from Clontech (mixed population of men aged 14-60). Two different batches were tested. The cDNA was prepared from this RNA, either, by reverse transcription (SuperScript III Reverse Transcriptase cat. no. 18080-093) or by One-Step RT-PCR (Qiagen cat. no. 210212).
Human total spermatozoa RNA (see Ethics statement) was prepared with the TriPure Isolation reagent (Roche), and/or RNeasy silica-membrane spin-columns (Qiagen) following the manufacturer instructions.
Cryptorchidic testis was immediately frozen and reduced to a homogenous powder (mortar) in liquid nitrogen. Total RNA was extracted with TriPure Isolation reagent, followed by further purification in RNeasy silica-membrane spin-columns (Qiagen), as before.
The procedure used for electrophoresis, northern blot, and preparation of DNA probes has been previously described [20 (link)].
Human VEGFR-1 kinase-domain primers (Table 1 ) were used to obtain a probe that hybridized with the full-length VEGFR-1 mRNA and any VEGFR-1 containing the whole or part of the kinase domain and C-tail.
Human total spermatozoa RNA (see Ethics statement) was prepared with the TriPure Isolation reagent (Roche), and/or RNeasy silica-membrane spin-columns (Qiagen) following the manufacturer instructions.
Cryptorchidic testis was immediately frozen and reduced to a homogenous powder (mortar) in liquid nitrogen. Total RNA was extracted with TriPure Isolation reagent, followed by further purification in RNeasy silica-membrane spin-columns (Qiagen), as before.
The procedure used for electrophoresis, northern blot, and preparation of DNA probes has been previously described [20 (link)].
Human VEGFR-1 kinase-domain primers (