Supplementary Figure 11: Identification of the key mRNA targets of miR-137.
(a) To identify the direct mRNA targets of miR-137, we overlapped the predicted miR-137 targets with those DE proteins and identified 41 upregulated miR-137 predicted targets. The targets ranked from high to low in average log2 fold change of two replicates. (b) qPCR measured luciferase mRNA levels in two of our reporter gene assays (Ptpn2 and Pde10a). The decreased luciferase level was regulated by the overexpressed miR-137 rather than altered luciferase mRNA level, demonstrating by the little-changed expression level of luciferase. All data shown are means ± s.e.m. n = 9 independent samples. tPtpn2 = 0.954, P = 0.3543; tPde10a = 0.3656, P = 0.7195. Unpaired Two-Tailed t- test; n.s., nonsignificant. (c) Western blot was performed to validate the result of the proteomics analysis. In 12- day-old mice cortex tissue, protein expression of the most upregulated miR-137 target was examined (n = 3 mice per group). GAPDH (AM4300; Thermo Fisher) was used for loading controls. ImageJ software quantified the band intensities. All data shown are means ± s.e.m. tPtpn2 = 2.496, P = 0.1300; tSatb2 = 0.985, P = 0.3804; tPde10a = 4.217, P = 0.0135. Unpaired Two-Tailed t- test; n.s., nonsignificant; *, P < 0.05.
