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. 2003 Sep;163(3):1177–1184. doi: 10.1016/S0002-9440(10)63477-3

Figure 2.

Figure 2.

Detection of Ikaros isoforms in human pituitary tumors. a: RT-PCR detection of Ik isoforms. RNA from primary human pituitary adenomas was reverse-transcribed and amplified using nested PCR primers as shown in Figure 1 ▶ . The expected product sizes are approximately as follows: Ik1 to Ik3, 1.3 to 1.5 kb; Ik4 to Ik8, 0.9 kb. Pituitary tumors (T1 to T6) express Ik1 to Ik3 and also yield a band of 0.9 kb; sample T1 submitted for PCR without reverse transcription (−) yields no product. The identity of all PCR products was confirmed by DNA sequencing; the 0.9-kb bands were verified by sequencing to be Ik6. The products of the RT reactions amplified with PGK-1 primers yield the expected 338-bp product, confirming intact quality of RNA (bottom). b: Ik isoform expression in human pituitary tumors by Western blotting. Lysates from GH4 cells and primary human pituitary tumors (T) were separated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotted with an Ik antibody that recognizes the C-tail that is common to all Ik isoforms. Note the expression of Ik1 at ∼57 kd and a smaller protein consistent with the size of Ik6 (∼36 kd) identified in several pituitary tumors. Immunoblotting with an anti-actin antibody confirms similar protein loading (bottom).