Fig 2: The regulation of circ_0124644 silencing on hypoxia-induced cardiomyocytes injury. (A) AC16 cells were transfected with sh-NC or sh-circ_0124644. The expression of circ_0124644 was measured by qRT-PCR. (B–I) AC16 cells were transfected with sh-NC or sh-circ_0124644, followed by treated with hypoxia. AC16 cells treated with normoxia were used as control. (B) CCK8 assay was used to examine cell viability. (C,D) Flow cytometry was performed to detect cell cycle distribution and cell apoptosis. (E) The protein levels of CyclinD1 and Cleaved-casp3 were determined using WB analysis. (F–I) Corresponding Assay Kits were used to assess the levels of LDH, MDA, SOD, and CAT. **P < 0.01, ***P < 0.001.

Fig 3: Klotho deficiency inhibited OGG1 aggravating podocytes injury in STZ-induced mice. (a) Representative western and summarized data showing the relative protein levels of OGG1 in the kidney from WT Vehicle, WT STZ, KL+/- Vehicle and KL+/- STZ. (b) Immunofluorescence staining showing the expression of OGG1 as well as podocyte membrane marker Synapotopodin in WT Vehicle, WT STZ, KL+/- Vehicle and KL+/- STZ. Scale bar, 20 µm. (c) Effect of Klotho on ROS generation in HG-induced podocytes. (d) Representative western blot gel documents and summarized data showing the relative protein level of Podocin, WT1, OGG1 and Caspase-3 p17 subunit in podocytes. *P<0.05; **P<0.01; ***P<0.001.

Fig 4: The regulation of miR-590-3p overexpression on cardiomyocytes injury induced by hypoxia. AC16 cells were transfected with miR-NC or miR-590-3p mimic, followed by treated with hypoxia. Normoxia-treated AC16 cells were used as control. (A) Cell viability was determined using CCK8 assay. (B,C) Cell cycle distribution and cell apoptosis were detected by flow cytometry. (D) WB analysis was performed to evaluate the protein levels of CyclinD1 and Cleaved-casp3. (E–H) The levels of LDH, MDA, SOD, and CAT were determined using corresponding Assay Kits. **P < 0.01, ***P < 0.001.

Fig 5: Klotho deficiency (KL+/-) promoted podocyte injury in STZ-induced mice. (a) Immunohistochemistry staining showing the expression of podocyte nuclear marker WT1 and Caspase-3 p17 subunit related to apoptosis in WT Vehicle, WT STZ, KL+/- Vehicle and KL+/- STZ. Scale bar, red 50 µm, black 20 µm. (b) Immunofluorescence staining showing the expression of podocyte membrane marker Nephrin in WT Vehicle, WT STZ, KL+/- Vehicle and KL+/- STZ. Scale bar, white 20 µm. (c) Representative photomicrographs and quantifications of mean glomerular basement membrane (GBM) thickness (yellow arrow), mean foot process width (red arrow), and the number of foot processes in different groups of mice by transmission electron microscopy (TEM) analyses. Scale bar, 200 nm. (d) Representative western blot gel documents and summarized data showing the relative protein level of Nephrin, PRDX2 and Bax in the kidney from WT Vehicle, WT STZ, KL+/- Vehicle and KL+/- STZ. *P<0.05; **P<0.01; ***P<0.001.