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. 2018 Apr 26;10(6):1807–1820. doi: 10.1016/j.stemcr.2018.03.022

Figure 4.

Figure 4

METTL8 Binds to Mapkbp1 mRNA

(A) Schematic illustration of PAR-CLIP assay.

(B) E14 cells were harvested and subjected to cell fractionation. Each fraction was collected and analyzed by western blot.

(C) HEK293T cells were transfected by the indicated plasmids. After 48 hr cells were harvested and subjected to RIP with flag antibody. Total RNAs were extracted followed by real-time PCR analysis. Actin was used as an internal control. Data are shown as the mean ± SD from three independent experiments. ∗p < 0.05.

(D) E14 cells were harvested and subjected to RIP with METTL8 antibody or isotype-matched immunoglobulin G (IgG). Total RNAs were extracted and relative level of Mapkbp1 mRNA was analyzed by quantitative real-time PCR. Actin was used as an internal control. Data are shown as the mean ± SD from three independent experiments. ∗p < 0.05.

(E) E14 cells were harvested and cell lysates were then incubated with in-vitro-synthesized biotin-labeled sense or antisense Mapkbp1 mRNA probes for biotin pull-down assay, followed by western blot analysis.