FIG. 2.

Analysis of HSV-1 2.0- and 1.5-kb LATs in transgenic-mouse tissues. (A) Northern blot analysis. Total RNAs were obtained from mouse tissues. The positions of RNA markers (in kilobases) are indicated on the right. The locations of the 1.5- and 2.0-kb LATs are indicated on the left. The DNA probe used was the BstEII-BstEII DNA fragment (Fig. 1A, II). (I) RNA (0.5 μg) was loaded in each lane. Lanes 1 to 7, transgenic-mouse tissues; lanes 8 to 11, normal-mouse tissues. To verify that equal amounts of RNA were loaded, the same blotted membrane was rehybridized with a 5′-end-labeled single-stranded DNA probe specific for the 18S rRNA, as described before (20) (data not shown). (II) RNA(10 μg) was loaded in each lane. (B) RT-PCR analysis. Shown are RT-PCR products from total RNAs extracted from various tissues (lanes 1, 3, 5, 7, 9, 11, and 13). A 254-bp RT-PCR DNA product represents the 1.5-kb spliced RNA, while a larger, 814-bp product represents the unspliced 2.0-kb transcript (18). For each sample, a control PCR was performed with no RT enzyme added (−) (lanes 2, 4, 6, 8, 10, 12, and 14). Lane 15, control RT-PCR with sample without RNA (−RNA). Lane 16, RT-PCR of RNA extracted from TG of normal mouse. Lane 17, RT-PCR of RNA extracted from LAT-expressing neuronal cells (20). Lane 18, PCR of the pLAT plasmid containing the LAT gene. Lane M, 100-bp DNA ladder (Promega). The PCR products were analyzed on a 1.8% agarose gel stained with ethidium bromide.