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. 2009 Jan 5;29(6):1442–1451. doi: 10.1128/MCB.01689-08

FIG. 2.

FIG. 2.

Exon 6A in the human hnRNP L gene is an NMD target. (A) HnRNP L exon 6A inclusion is increased after cycloheximide treatment. HeLa cells were treated with cycloheximide over 3 h, and total RNA was prepared after 0, 0.5, 1, and 3 h (as indicated) and analyzed by semiquantitative RT-PCR for exon 6A inclusion (top). Quantitation of exon 6A inclusion is given below the lanes (in percentages). The panels below show control RT-PCRs for β-actin mRNA (middle), and for alternative splicing of another hnRNP L target gene, TJP1 (bottom; the band marked by the asterisk represents a PCR artifact). RT-PCR products are schematically represented on the right. (B and C) hnRNP L exon 6A inclusion is increased after UPF1 knockdown (ΔUPF1). UPF1 expression was downregulated in HeLa cells by RNAi, with a luciferase knockdown (Δluc) as a control (see panel B for Western blot analysis of UPF1 and control γ-tubulin protein levels). (C) Total RNA after knockdowns of UPF1 (ΔUPF1) and control luciferase (Δluc) was analyzed for exon 6A inclusion by RT-PCR (RT-PCR products schematically represented on the right). Quantitation of hnRNP L exon 6A inclusion is given below the lanes (in percentages). M, DNA size markers.