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. 2022 Mar 14;13(5):1664–1678. doi: 10.7150/jca.57691

Figure 4.

Figure 4

MiR-130b-3p could bind with circAPSM and mediated the function of GBM cells. a, b: MiR-130b-3p inhibitor treatment up-regulated the expression of circAPSM while mimic treatment down-regulated circASPM expression as measured by qPCR. c: The predicted binding site between circASPM and miR-130b-3p. d, e: The expression of miR-130b-3p was down-regulated after circAPSM overexpression while circASPM knockdown up-regulated miR-130b-3p expression as measured by qPCR. f, g, h, i: The luciferase reporter assays showed that miR-130b-3p mimic or inhibitor altered the luciferase promoter activities of circASPM. j: FISH assays detected the subcellular localization of circASPM and miR-130b-3p. k, l: CircASPM and miR-130b-3p were effectively pulled down by anti-AGO2 antibodies compared to IgG, and both enriched after miR-130b-3p mimic treatment in GSC18. m: The RNA pull-down assays demonstrated circASPM can sponge miR-130b-3p. n, o: MTS assays showed that circASPM transfection of overexpression plasmids or si-circASPM affected GSCs viability and was reversed by miR-130b-3p mimic or inhibitor treatment, respectively. p: The EDU assay showed that circASPM transfection of overexpression plasmids or si-circASPM affected GSCs proliferation capacity and was reversed by miR-130b-3p mimic or inhibitor treatment, respectively. Scale bar = 50 μm. q, r: In the neurosphere formation assays and extreme limiting dilution assays, circASPM transfection of overexpression plasmids or si-circASPM affected neurosphere growth capacity in GSCs and was reversed by miR-130b-3p mimic or inhibitor treatment, respectively. Scale bar = 50 μm. All data are shown as the mean ± SD (three independent experiments). *P < 0.05; **P < 0.01; ***P < 0.001.