Fig 1: miR-370-3p overexpression-mediated protective effects in hypoxia-induced cardiomyocytes are partly counteracted by the introduction of PDE4D plasmid. (a) The transfection efficiency of PDE4D plasmid in AC16 cells was assessed by Western blot assay. (b–l) AC16 cells were divided into the following four groups: hypoxia+miR-NC, hypoxia+miR-370-3p, hypoxia+miR-370-3p+vector, and hypoxia+miR-370-3p+PDE4D. (b) CCK8 assay was conducted to assess cell viability. (c) Edu assay was performed to analyze the proliferation ability of AC16 cells. (d) Cell migration ability was assessed by Transwell migration assay. (e) The apoptosis of AC16 cells was analyzed by flow cytometry. (f, g) Cell oxidative status was analyzed using the matched kits. (h) The release of IL-6 and TNF-a was analyzed by ELISA. (i–k) The uptake of glucose and the production of lactate and ATP were analyzed using their matched colorimetric assay kits. (l) The expression of HK2 and LDHA was analyzed by Western blot assay. *P < 0.05.
Fig 2: The role of the circ_0023461/miR-370-3p/PDE4D axis on hypoxia-induced AC16 cells.
Fig 3: miR-370-3p directly binds to PDE4D in cardiomyocytes. (a) The interacted mRNAs of miR-370-3p were predicted by starBase database, and PDE4D was one of the candidate targets of miR-370-3p. (b, c) The binding relationship between miR-370-3p and PDE4D was verified by dual-luciferase reporter assay and RNA pull-down assay. (d, e) The effect of miR-370-3p overexpression on the mRNA and protein expression of PDE4D in AC16 cells was analyzed by RT-qPCR and Western blot assay. (f, g) The mRNA and protein expression of PDE4D was determined in the plasma samples of AMI patients and normal volunteers by RT-qPCR and Western blot assay. (h, i) The mRNA and protein levels of PDE4D were examined in hypoxia-treated AC16 cells by RT-qPCR and Western blot assay. *P < 0.05.
Fig 4: PDE4D is regulated by the circ_0023461/miR-370-3p axis in AC16 cells. (a, b) The mRNA and protein expression of PDE4D was examined in AC16 cells by RT-qPCR and Western blot assay in the following four groups: hypoxia+si-NC, hypoxia+si-circ_0023461, hypoxia+si-circ_0023461+anti-miR-NC, and hypoxia+si-circ_0023461+anti-miR-370-3p. *P < 0.05.
Supplier Page from Abcam for Anti-PDE4D antibody [EPR11823-31] - BSA and Azide free