Fig. 5

Validation of the targeting relationship between Crnde and miR-181a-5p and that between miR-181a-5p and Pcyox1l. A, RNA-FISH assay for detection of the co-localization of Crnde and miR-181a-5p in the nucleus and quantitative statistical analysis results. B, The binding site and mutation site of Crnde and miR-181a-5p. C, Dual luciferase reporter gene assay to verify the binding of Crnde to miR-181a-5p. D, The direct binding between Crnde and miR-181a-5p detected by RNA pull-down assay. E, The binding site and mutation site of Pcyox1l and miR-181a-5p. F, Dual luciferase reporter gene assay to verify the binding of Pcyox1l mRNA to miR-181a-5p. G, The interaction relationship between Pcyox1l mRNA to miR-181a-5p as detected by RIP assay. H, Pcyox1l and miR-181a-5p expression in the DVT mice in response to Crnde knockdown or miR-181a-5p agomir measured by RT-qPCR. I & J, Images (I) and quantitation (J) of the Pcyox1l expression in the DVT mice in response to Crnde knockdown or miR-181a-5p agomir determined by Western blot. K & L, Images (K) and quantitation (L) of Pcyox1l protein expression in the DVT mice in response to Crnde knockdown or miR-181a-5p agomir determined by immunohistochemistry. *** p < 0.001 vs. the mimic-NC, NC probe, sh-NC or agomir NC group. n = 6. All cell experiments were independently repeated three times