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. 2022 Apr 22;13(4):397. doi: 10.1038/s41419-022-04854-1

Fig. 5. Itch is directly targeted by miR-144.

Fig. 5

A Predicted binding sites of miR-144 in the 3ʹUTR of Itch. The miR-144-3p seed region is shown. The mutated sequence (mt) was changed to lower-case letters. B H1299 cells were transfected with the indicated expression constructs. Luciferase activities were measured. C The levels of miR-144 expression were measured by qRT–PCR. D, E miR-144 negatively regulates Itch protein levels. Plasmids expressing control-miRNA or miR-144 were transfected into H1299 (D) or SCC9 (E) cells as shown. The levels of endogenous p63 and Itch protein were detected by Western blotting. Actin was used as a loading control. F The levels of Itch mRNA were measured by qRT–PCR in H1299 and SCC9 cells. G The levels of miR-144 expression were detected by qRT–PCR in H1299 and SCC9 cells. All experiments were performed in triplicate. *P < 0.05, **P < 0.01, ***P < 0.001.