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. 2022 May 18;12:876090. doi: 10.3389/fonc.2022.876090

Figure 4.

Figure 4

HAGLROS regulates SPRR1B expression by sponging miR-330-5p. (A) The specific binding sites among HAGLROS, miR-330-5p and SPRR1B were predicted by the Lncbase database and the starBase database. (B, C) MiR-330-5p expression was examined in bladder tissues and cells using qRT–PCR assays. (D) The changes of miR-330-5p expression were determined using qRT–PCR assays. (E) FISH assay was used to confirm the subcellular colocalization between HAGLROS and miR-330-5p in BC cells (magnification, x 400). HAGLROS (Cy3, red), miR-330-5p (FAM, green), and cell nuclei (DAPI, blue). (F) FISH assay was used to confirm the subcellular colocalization between miR-330-5p and SPRR1B in BC cells (magnification, x 400). SPRR1B (Cy3, red), miR-330-5p (FAM, green), cell nuclei (DAPI, blue). (G) Wild-type or mutant HAGLROS was cotransfected with mimic-NC or miR-330-5p mimic into HEK-293T cells, and the relative luciferase activities were measured. (H) Wild-type or mutant SPRR1B was cotransfected with mimic-NC or miR-330-5p mimic into HEK-293T cells, and the relative luciferase activities were determined. Each experiment was repeated at least thrice. Ns, not significant. **P < 0.01, ***P < 0.001, ****P < 0.0001.