Inclusion of hnRNP L Alternative Exon 7 Is Associated with Good Prognosis and Inhibited by Oncogene SRSF3 in Head and Neck Squamous Cell Carcinoma

Figure 4

The inhibition of SRSF3 on the inclusion of hnRNP L exon 7 was evaluated by CHX treatment. (a) CHX was used to block NMD. Cells were treated with CHX or DMSO for 5 hours. The alternative splicing of exon 7 was analyzed by RT-PCR. L/S ratio represents the ratio of exon 7 inclusion versus exclusion isoform. (b-c) CAL 27 or SCC-9 cells were treated with anti-SRSF3 siRNA (si#2) or nonspecific control siRNA (NC), followed by CHX treatment. Knockdown efficiency of SRSF3 was analyzed by Western blot (b). Alternative splicing of hnRNP L exon 7 was analyzed by RT-PCR (c). ◄: hybridization product of both long and short isoforms. (d) The histogram summarized the effects of SRSF3 knockdown on the alternative splicing of hnRNP L exon 7 in CAL 27 and SCC-9 cells. Data are the means ± SD, n = 3. : , : . (e) CAL 27 or SCC-9 cells were transfected with SRSF3 expression plasmid or vector control plasmid, followed by CHX treatment. The alternative splicing of exon 7 was analyzed by RT-PCR. Overexpression of SRSF3 was confirmed by Western blot. (f) The histogram summarized the effects of SRSF3 overexpression on the alternative splicing of hnRNP L exon 7 in CAL 27 and SCC-9 cells. Data are the means ± SD, n = 3.

(a)

(b)

(c)

(d)

(e)

(f)