Northern blotting analysis for the expression of 5 novel cytokine-responsive genes in resting and TNF-–activated HUVEC. Northern blotting analysis of 10 μg HUVEC total RNA with probes from DD-fragments or corresponding EST-clones was performed as described in Materials and Methods. Time periods of continuous stimulation by TNF- are indicated, and GAPDH analysis is given as control for equal loading. The approximate length of the transcripts was determined from the position of 28S, 18S, and 5S ribosomal RNA at the following: CA2_1: an abundant band of 5.2 kb and two minor bands of 6 and 4.4 kb, respectively; GG10_2: 2.8 kb; GG2_1: 1.8 kb; CG12_1: 2.3 kb; and AG8_1: 3.2 kb. A detailed analysis of these 5 transcripts is given in the text and in Fig 4. In the case of CA2_1, identical patterns and intensities were found with radiolabelled probes from bases 639-1032 (DD-fragment) and 2331-3317 (hIAP-1) from the full-length sequence. The other probes that were used represent the following parts of the full-length sequences: GG10_2: 2697-2883; GG2_1: 670-1882; CG12_1, 1690-2298; AG8_1: 2880-3289; and GAPDH: 360-1070 (GB: M33197).