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. Author manuscript; available in PMC: 2016 Nov 3.
Published in final edited form as: Int J Dev Biol. 2016;60(7-8-9):315–320. doi: 10.1387/ijdb.160161ds

Figure 5. Sperm Chromatin transcription in oocyte and egg extracts.

Figure 5

A. Sperm chromatin was incubated in oocyte or egg extracts in the presence of 32P-α-ATP. Sperm was pre-incubated with TBP protein as indicated. The reaction products were stopped and RNA extracted with TRIzol. Precipitated RNA was directly counted in a scintillation counter and resulting transcription radioactivity was scaled to the oocyte sperm control.

B. Sperm chromatin was incubated in egg extract either without or pre-incubated with TBP protein as indicated. The reaction product chromatin was isolated after 0, 15, 30, 60 or 90 minutes through a sucrose cushion and pelleted proteins were immunoblotted for PCNA and histone H2A.

C. Sperm chromatin was incubated in oocyte or egg extracts. The reaction product chromatin was isolated after 0, 15, 30, 60 or 90 minutes through a sucrose cushion and pelleted proteins were immunoblotted for RNA Pol II, Pol II S5ph, and H2A as indicated. A total Coomassie stained gel of the isolated products is also shown at the bottom.