Figure 2.

Confirmation of α subunit gene inactivations in the mutant ES cell lines. (A) Western blot analysis of membrane protein from undifferentiated WT and α-null cells. (B) Northern blot analysis of total RNA prepared from 7- to 10-day-old differentiated cells and probed with a 1.1-kb NcoI–EcoRI fragment of α_o cDNA containing exons 3 through 8 revealed the presence of multiple transcripts in WT cells but none in the α_o-null cells. Non-specific binding to 18S and 28S RNA was observed in all lanes. (C) RT-PCR of total RNA was used to assess α_q expression and expression of the highly related α₁₁ gene product. An α_q product of the predicted size of 655 bp was amplified only in WT cells while the control amplification for α₁₁ detected a product (predicted size of 589 bp) in both WT and the α_q-null cells.