Fig 1: Expression of lncRNA EBLN3P, TMPRSS3 and NT-3 in the cochlear SGNs of deafness models. (A) Fluorescence in situ hybridization was performed to observe the expression of lncRNA EBLN3P in the cochlear SGNs of deafness models. (B and C) IHC and quantitative analysis were performed to observe the expression of TMPRSS3 in the cochlear SGNs of deafness models. (D and E) IHC and quantitative analysis were performed to observe the expression of NT-3, an important growth factor that helps stimulate and control neurogenesis, in the cochlear SGNs models of deafness. **P<0.01 (Student's t-test) vs. model. lncRNA, long non-coding RNA; SGN, spiral ganglion neuron; TMPRSS3, transmembrane protease, serine 3; NT-3, neurotrophin-3; IHC, immunohistochemistry.
Fig 2: Schematic model of lncRNA EBLN3P functioning as a competing endogenous RNA for miR-204-5p and regulating the expression of TMPRSS3. miR-204-5p could bind to the 3'-untranslated region of TMPRSS3 to inhibit the expression of TMPRSS3. lncRNA EBLN3P could bind to miR-204-5p at two sites, '314-322' and '973-981', in the 1,200-nt gene fragment at the 5'-end of EBLN3P, and block the regulatory function of miR-204-5p on TMPRSS3 expression. lncRNA, long non-coding RNA; miR, microRNA; TMPRSS3, transmembrane protease, serine 3.
Fig 3: lncRNA EBLN3P regulates the expression of TMPRSS3 through miR-204-5p in normal SGNs. (A) RT-qPCR was performed to detect the effects of lncRNA EBLN3P knockdown on the expression of EBLN3P in primary normal SGNs. (B) Luciferase activity was evaluated to observe the effect of lncRNA EBLN3P knockdown on the psiCHECK-2/TMPRSS3 wild-type 3'-UTR, psiCHECK-2/TMPRSS3 mutated 3'-UTR and empty 3'-UTR vectors (control). (C) RT-qPCR was performed to detect the effects of lncRNA EBLN3P knockdown on the mRNA expression of TMPRSS3 in primary normal SGNs. (D) Western blot analysis was performed to detect the effects of lncRNA EBLN3P knockdown on the protein expression of TMPRSS3 in primary normal SGNs. (E) RT-qPCR was performed to detect the effects of lncRNA EBLN3P overexpression on the expression of EBLN3P in primary normal SGNs. (F) Normal SGNs were treated with over-NC, over-lncRNA EBLN3P or anti-miR-204-5p + over-lncRNA EBLN3P, and the luciferase activity was evaluated. (G-I) Normal SGNs were treated with control over-NC, over-lncRNA EBLN3P or anti-miR-204-5p + over-lncRNA EBLN3P. RT-qPCR and western blot analysis was performed to detect the mRNA and protein expression of TMPRSS3 in primary normal SGNs. **P<0.01 (Student's t-test) vs. NCs in panels A, B, C, D and E. **P<0.01 (ANOVA with Tukey's post hoc test) in panels F, G and I. lncRNA, long non-coding RNA; SGN, spiral ganglion neuron; TMPRSS3, transmembrane protease, serine 3; UTR, untranslated region; NC, negative control; miR, microRNA.
Supplier Page from Abcam for Anti-TMPRSS3 antibody [EPR7048]