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. 2024 May 29;15:4549. doi: 10.1038/s41467-024-48740-0

Fig. 2. miR-199b targets SLC transporters EAAT2, SNAT2, and MCT2.

Fig. 2

a Predicted miR-199b binding sites in the 3′UTR of human SLC1A2 (EAAT2), SLC38A2 (SNAT2), and SLC16A7 (MCT2). The corresponding wild-type (WT) sequences are shown. For each gene, a 3′UTR fragment encompassing both miR-199b binding sites was cloned into a reporter plasmid. Highlighted sequences were mutated to destroy site I (mut1), site II (mut2), or both (mut1 + 2) in the reporter construct. Responsiveness of the reporters to miR-199b was determined in MDA-MB-231 cells overexpressing miR-199b or control (n = 3 biological replicates). b RT-qPCR-determined RNA levels of EAAT2 in astrocytes (NHA) and levels of SNAT2 and MCT2 in neurons (differentiated SH-SY5Y) upon transfection with miR-199b or control mimic. Data were normalized to 18S rRNA (n = 3 biological replicates). c Western blot showing levels of indicated proteins in astrocytes and neurons upon miRNA mimic transfection. GAPDH was used as a loading control. In bar graphs, values are shown as mean ± SD. Unpaired two-tailed t-test was used in a and b. P values are indicated. Source data are provided as a Source Data file.