Cycloheximide dependence of TNF-α gene transcription in activated human monocytes

Effect of tumor necrosis factor-α on thein vitro synthesis of interleukin-8 by human monocytes and lymphocytes

“…The DNA probes were labeled with a-~2P d-ATP by primer elongation with Klenov's fragment of E. coli DNA polymerase 1 to attain the specific activity of about 10 9 decays/gg [12]. Hybridization with labeled probes and membrane washing were performed as previously [11]. The results were analyzed using Student's t test.…”

“…The results were analyzed using Student's t test. and interleukin-l(3 (IL-1) as described previously [11]. The total RNA was isolated from the cells as described elsewhere [2] after cell lysis with 4 M guanidine isothiocyanate (Fluka).…”

The Role of 3′ Poly(A) Tail Metabolism in Tumor Necrosis Factor-α Regulation

“…This possibility was addressed by analyzing TNF-␣ mRNA content and size after treating RAW cells with cycloheximide, an agent that induces TNF-␣ transcription (30) and increases TNF-␣ mRNA level (31). Cycloheximide (CHX) increased TNF-␣ transcription rate 80% (Fig.…”