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Fig. 2 | Journal of Neuroinflammation

Fig. 2

From: miR-17-92 facilitates neuronal differentiation of transplanted neural stem/precursor cells under neuroinflammatory conditions

Fig. 2

miR-17-92 cluster members target multiple proteins in JAK2/STAT3 pathway. a The schematic genomic organization of the miR-17-92 cluster on the mouse chromosome 14 (Chr.14). b Sequence alignment of mature miR-19a, miR-19b, miR-17, and miR-20a revealed their seed sequences that were reverse complementary to the seed-matched sequence within the 3′ UTR of mouse CNTFR or GP130, respectively. The mutated sequences in the 3′ UTRs of CNTFR (mut CNTFR) and GP130 (mut1 GP130, mut2 GP130) are indicated. c, d Luciferase activity was measured 24 h after transfecting HEK 293T cells. Reporter plasmids with the wild-type (wt CNTFR, wt GP130) or mutated (mut CNTFR, mut1 GP130, or mut2 GP130) 3′UTR of CNTFR or GP130 were transfected either alone (vector) or with the expression vector pcDNA3.1-overexpressing miR-17-92 cluster (miR-17-92) or empty pcDNA 3.1 (con). Values are means ± SEM. Asterisks indicate a statistically significant difference compared with the control (*P < 0.05, one-way ANOVA followed by Tukey’s test, n = 3 independent experiments). e Quantitative real-time PCR was used to determine the efficiency of overexpressing miR-17-92 cluster members using lentivirus (miR-17-92). Values are means ± SEM (n = 3 independent experiments). f Western blot analysis of GP130, CNTFR, JAK2, and STAT3 protein expressions in NSCs 72 h after infecting them with lentivirus overexpressing miR-17-92 cluster (miR-17-92) or control lentivirus (CON). g Quantifications for the protein expression levels from the experiment shown in f. Values are means ± SEM. Asterisks indicate a statistically significant difference compared with the control group (*P < 0.05, unpaired two-tailed t test, n = 3 independent experiments). h Schematic representation of multiple proteins regulated by miR-17-92 cluster in JAK2/STAT3 signaling pathway

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