Fig 2: Luciferase assays confirmed the 3, UTR sites of MAP3K2 and MAP2K4 targeted by miR-512-3p and miR-519a-2-5p. (A) The predicted binding site sequence in the 3′UTR of MAP3K2 and MAP2K4 and the Corresponding mutated site. (B) Dual-luciferase assays showed the relative luciferase activities (Firefly/Renilla) among the HEK-293T cells co-transfected with miR-512-3p (left), miR-519a-2-5p (right) or control mimics and luciferase wild-type or mutant reporters. The statistics analysis was performed with the one-way ANOVA test. “***” and “****” represented the p-value < 0.001 and 0.0001, respectively.

Fig 3: The schematic diagram showed that the miR-512-3p and miR-519a-2-5p promoted hepatocellular carcinoma progression by co-targeting the MAP3K2 and MAP2K4.

Fig 4: Cellular localization of map3k2 and csnk1e in frozen sections of naïve lamprey brains and spinal cords. (A) Western blot of proteins prepared from lamprey brains (Br) or spinal cords (SC), probed with antibodies against map3k2 and csnk1e. A single band at approximately 34 and 44 KDa was labeled with antibodies to map3k2 and csnk1e, respectively, in either brains or spinal cords. (B,C) Chromogenic IHC of coronal brain sections stained for map3k2 and csnk1e, respectively. Rostral is up. The two insets at the bottom right of (B,C) are enlarged images of the I1 neurons framed by the respective white boxes. Positive immunostaining is in brown; counterstaining by hematoxylin is in light blue. (D) Spinal cord coronal sections immuno-fluorescently triple stained for map3k2 (red), csnk1e (green) and DAPI (blue). The three asterisks (*) are inside faintly stained uninjured axons. CC: central canal. (E1–E3) are monochromatic enlarged micrographs from the framed region in (D). (E4) is (E1–E3) superimposed, showing local neurons (1–3) and glial cells (4).

Fig 5: Volcano maps showing the consistently greater expressions of map3k2 and csnk1e in G tips than in R tips or S tips: (A) Volcano plot of genes that were differentially expressed in G vs. R tips. Blue dots: 513 genes that were comparatively upregulated (right side) or downregulated (left side) in G tips compared with R tips (p values < 0.01). Red and Green dots represent the genes map3k2 and csnk1e, respectively. (B) Volcano plot of 151 genes that were differentially expressed in G tips vs. S tips (p < 0.01).