Stem Cells International / 2017 / Article / Fig 4

Research Article

The DEAD-Box RNA Helicase DDX3 Interacts with m6A RNA Demethylase ALKBH5

Figure 4

DDX3 modulated m6A RNA demethylation. (a) DDX3 knockdown in HEK293T cells, relative DDX3 mRNA (upper panel, qPCR), and protein levels (lower panel, Western blots) are shown. (b) ALKBH5 knockdown in HEK293T cells, ALKBH5 mRNA (upper panel, qPCR), and protein levels (lower panel, Western blots) are shown. (c) Left panel: dot-blot analyses of m6A levels of isolated total RNA, mRNA, and miRNA from ALKBH5 knockdown, DDX3 knockdown, and control (NC, siRNA with scrambled sequences) cells. Right panel: methylene blue staining showing equal RNA loading. (d) IP of endogenous DDX3 could co-IP endogenous AGO2, and IP of endogenous AGO2 could co-IP endogenous DDX3 in HEK293T cells. (e) Quantification of cell proliferation (MTT assay) after knockdown of DDX3 or ALKBH5 in HEK293T cells. (f) Quantification of cell proliferation (MTT assay) after knockdown of DDX3 or ALKBH5 in HeLa cells. Dot blots, IP, and co-IP were performed in triplicates, and representative results were shown. Scramble siRNA with scrambled sequences. , , and . values were determined with two-tailed Student’s t-test. Error bars represent standard deviation (SD).
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