Fig. 2: DCP is targeted by WFDC21P, which is elevated by miR-4293. | Cell Death & Disease

Fig. 2: DCP is targeted by WFDC21P, which is elevated by miR-4293.

From: miR-4293 upregulates lncRNA WFDC21P by suppressing mRNA-decapping enzyme 2 to promote lung carcinoma proliferation

Fig. 2

a TargetScan analysis predicted that the DCP2 mRNA-3′-UTR is targeted by miR-4293. We synthesized a mutant miR-4293 (Mu-4293) and cloned DCP2 mRNA-3’-UTR and mu-DCP2–3’-UTR sequences. b Analysis of miR-4293 regulating luciferase expression of reporter plasmids containing DC2–3’UTR (right panel) or mu-DCP2–3’UTR (left panel). Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; Student’s t-test. c Analysis of ASO-4293 regulating luciferase expression of reporter plasmids containing DC2–3’UTR (right panel) or mu-DCP2–3’UTR (left panel). Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; Student’s t-test. d Western blot detection of DCP2 in miR-4293-, ASO-4293-, or scramble-treated A549 cells. e Western blot detection of DCP2 in siRNA-DCP2–1-, siRNA-DCP2–2-, or scramble-treated A549 cells and H129 cells. f qRT-PCR analysis of WFDC21P level in pcDNA-WFDC21P + scramble-, si-DCP2-, si-DCP2 + pcDNA-WFDC21P-, or scramble-treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; ANOVA test. g Western blot detection of STAT3 phosphorylation in pcDNA-WFDC21P + scramble-, si-DCP2-, si-DCP2 + pcDNA-WFDC21P-, or scramble-treated A549 cells. h qRT-PCR analysis of DCP2 level in pcDNA or pcDNA-DCP2 -treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; Student’s t-test. i qRT-PCR analysis of WFDC21P level in 0 h, 4 h, 8 h, 12 h, 24 h, 48 h after transfection of pcDNA-DCP2. Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; ANOVA test. j RIP assay of the interaction between WFDC21P and DCP2. Upper panel, qRT-PCR analysis of WFDC21P pull-down by IgG or DCP2; lower panel, electrophoresis for WFDC21P pull-down by IgG or DCP2 after PCR amplification. Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; Student’s t-test. k qRT-PCR analysis of WFDC21P levels in miR-4293-, ASO-4293, Mu-4293 or scramble-treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; ANOVA test. l qRT-PCR analysis of WFDC21P levels in miR-4293+pcDNA-WFDC21P-, or scramble+ pcDNA-WFDC21P-treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. **p < 0.01; Student’s t-test. m Western blot detection of STAT3 phosphorylation in pcDNA-WFDC21P + scramble-, miR-4293-, miR-4293+pcDNA-WFDC21P-, or scramble-treated A549 cells. n Western blot detection of STAT3 phosphorylation in AG490 + pcDNA-, pcDNA-WFDC21P-, AG490 + pcDNA-WFDC21P-, or pcDNA treated A549 cells. o qRT-PCR analysis of miR-4293 level in AG490 + pcDNA-, pcDNA-WFDC21P-, AG490 + pcDNA-WFDC21P-, or pcDNA treated A549 cells. Data are expressed as mean ± SD for triplicate experiments. **p < 0.01, *P < 0.05; ANOVA test.

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