Fig 1: Targeting relationship between miR-21 and SPRY1. (a) miR-21 has the binding sites with SPRY1. (b) The luciferase activity in T/G HA-VSMC cells cotransfected with SPRY1-Wt, SPRY1-Mut, miR-21 mimic, and miR-NC was evaluated using dual-luciferase reporter assay. *P < 0.05.
Fig 2: Circ_0020093 positively regulated SPRY1 expression by targeting miR-23b. A The expression of SPRY1 in chondrocytes transfected with circ_0020093 or circ_0020093 + miR-23b was detected by Western blot, with pCD5-ciR or circ_0020093 + miR-con as the control. B The expression of SPRY1 in chondrocytes transfected with si-circ_0020093 or si-circ_0020093 + in-miR-23b was detected by Western blot, with si-con or si-circ_0020093 + in-miR-con as the control. *P < 0.05
Fig 3: MiR-23b targeted SPRY1 to aggravate IL-1ß-induced chondrocyte apoptosis and ECM degradation. IL-1ß-treated chondrocytes were transfected with in-miR-23b, in-miR-con, in-miR-23b + si-SPRY1, or in-miR-23b + si-con. In these transfected cells, A the expression of SPRY1 was measured by Western blot. B Cell apoptotic rate was analyzed by flow cytometry assay. C, D The expression of Aggrecan, COL2, MMP13, MMP3, and ADAMTS4 was detected by qRT-PCR and Western blot. *P < 0.05
Fig 4: Plasma levels of miR-21, SPRY1, and CX43. (a) The relative expression level of miR-21 in the plasma from CHD patients and healthy volunteers was detected using RT-qPCR analysis. (b) The mRNA expression of SPRY1 in the plasma from CHD patients and healthy volunteers was measured using RT-qPCR analysis. (c) The mRNA expression of CX43 in the plasma from CHD patients and healthy volunteers was determined using RT-qPCR analysis. (d, f) Western blot was used to assess the protein expression of SPRY1 protein in the plasma from CHD patients and healthy volunteers. (e, f) Western blot was used to examine the protein expression of CX43 in the plasma from CHD patients and healthy volunteers. (f) The representative images from western blot results. *P < 0.05.
Fig 5: Levels of miR-21, SPRY1, and CX43 in cells. (a) The relative expression of miR-21 was assessed by RT-qPCR analysis. (b) The relative mRNA expression of SPRY1 was determined by RT-qPCR analysis. (c) RT-qPCR analysis was utilized for CX43 mRNA expression measurement. (d, g) Western blot was performed to detect SPRY1 protein expression. (e, g) CX43 protein expression was determined by western blot. (f) The representative images from western blot results. *P < 0.05.
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