Fig 2: Elevated expressions of MMP-9 and MMP-13 were alleviated by GYY treatment in MCs under HG condition. (A) Untreated or GYY-treated mouse MCs in NG or HG condition were collected in Trizol for total RNA extraction, and semi-quantitative RT-PCR analyses were performed for MMP-9 and MMP-13 gene expression. (B) Protein was extracted from the untreated or GYY-treated mouse MCs in NG or HG condition and analyzed for the expression of MMP-9 and MMP-13 by Western blot. GAPDH was used as a loading control for all RT-PCR and immunoblot analyses. The bar graphs represent the mean fold change ± SD vs. NG. n = 3 independent experiments, * p < 0.05 vs. NG, † p < 0.05 vs. HG.

Fig 3: Elevated expressions of MMP-9 and MMP-13 were alleviated by GYY treatment in diabetic kidney. (A) Saline- or GYY-treated kidneys from WT and Akita mice were collected in Trizol for total RNA extraction, and semi-quantitative RT-PCR analyses were performed for MMP-9 and MMP-13 gene expression. (B) Protein was extracted from the saline- or GYY-treated kidneys from WT and Akita mice and analyzed for the expression of MMP-9 and MMP-13 by Western blot. GAPDH was used as a loading control for all RT-PCR and immunoblot analyses. The bar graphs represent the mean fold change ± SD vs. WT + Saline. n = 6/group, * p < 0.05 vs. WT + Saline, † p < 0.05 vs. Akita + Saline.

Fig 4: Circ_0058357 regulates the growth, invasion, migration, apoptosis and DDP sensitivity of H1299/DDP and A549/DDP cells in vitro. (a) qRT-PCR analysis of circ_0058357 in si-circ_0058357- or si-NC-transfected H1299/DDP and A549/DDP cells. (b) MTT assay presenting the IC50 for DDP in A549/DDP, A549, H1299/DDP, H1299 NSCLC cells. (c) H1299/DDP and A549/DDP cells were introduced with si-circ_0058357 or si-NC and exposed to various concentrations of DDP for 24 h, followed by the evaluation of the IC50 of DDP by MTT assay. (d)–(j) H1299/DDP and A549/DDP cells were introduced with si-circ_0058357 or si-NC for 48 h. (d) The viability of transfected H1299/DDP and A549/DDP cells was examined by MTT assay. (e) Representative images depicting a cell proliferation assay performed by the usage of EdU assay. (f) Representative pictures displaying a cell apoptosis assay performed by flow cytometry. (g) Representative images showing a cell invasion assay and transwell assay for cell invasion. (h) Representative pictures depicting a cell migration assay and cell migration by wound-healing assay. (i) and (j) The levels of CyclinD1, MMP9 and MDR1 in transfected H1299/DDP and A549/DDP cells were detected by western blot. **p < 0.01, ***p < 0.001, ****p < 0.0001

Fig 5: AdipoRon and leptin exert antagonizing effects on HIF-1α, CXCL1, VEGF-A, MMP-2 and MMP-9 protein levels. (A) Elisa assay of CXCL1 and VEGF-A; (B) representative images of immunoblotting of total MMP-2 and MMP-9 proteins and their quantitation. HUVEC endothelial cells were treated with AdipoRon (5 or 10 μg/mL), Leptin (125 ng/mL) and a combination of AdipoRon+leptin (5 μg/mL and 125 ng/mL, respectively) for 24 h. Untreated cells were used as negative control (NC); VEGF (10 ng/mL) was used as a positive control. Data are reported as mean ± Standard Deviation (SD) of three independent experiments were reported. * p < 0.05.