Fig 2: Microcystin (MC)-RR inhibits the expression of hypoxia inducible factor 1 subunit alpha (HIF-1α) and alters the level of matrix metalloproteinases (MMPs). Unilateral ureteral obstruction (UUO) mice and the cultured NRK-49F cells were treated as described in Figure 2 and Figure 4, respectively. (A,B) Expression of HIF-1α protein in the kidney tissues was detected by western blot. (C) Level of HIF-1α in NRK-49F cells was determined by western blot. (D,E) Expression levels of MMP-7, MMP-9, and MMP-13 in the kidney tissues of UUO mice were analyzed by western blot. (F) Deposition of collagen and elastic fibers in the kidney tissues of UUO mice with MC-RR treatment was evaluated by Sirius Red staining and Van Gieson staining (scale bar: 40 μm). *p < 0.05, **p < 0.01 determined by one-way ANOVA with S–N–K post hoc analysis, n = 5.

Fig 3: Notopterol abrogates RANKL-induced NFATc1 activation and associated downstream protein expression. (A,B) Representative confocal images (A) and quantification analysis (B) indicated that NFATc1 protein expression activated by RANKL was strikingly hampered by Notopterol. Scale bar = 200 μm. (C) RAW 264.7 cells transfected with an NFATc1 luciferase construct were pre-treated with indicated concentrations of Notopterol, followed by RANKL (50 ng/ml) stimulation for 24 h. NFATc1 luciferase activity was measured with a luciferase reporter assay system. (D) Representative images of Western blotting reflecting the effects of Notopterol on NFATc1, MMP9, c-Fos, CTSK induced by RANKL. (E–H) The ratios of band intensity of NFATc1 (E), MMP9 (F), c-Fos (G), CTSK (H) relative to β-actin were quantitatively determined. “−” means RANKL untreated; “+” means RANKL treated. Data are presented as mean ± SEM. All in vitro experiments were repeated three times with similar results. Significant differences between the control and treatment groups are shown as *p < 0.05, **p < 0.01 and ***p < 0.001.

Fig 4: miR-211-5p inhibitor alleviated the effects of TTN-AS1 knockdown on the expression of MMP-2 and MMP-9 in TNBC cells. Expression of MMP-2 and MMP-9 in (A) MDA-MB-453 and (B) MDA-MB-231 cells was evaluated using western blotting following transfection with shRNA-TTN-AS1-1 and/or miR-211-5p inhibitor. ***P<0.001 vs. shRNA-NC; #P<0.05, ##P<0.01 and ###P<0.001 vs. shRNA-TTN-AS1-1 + miR-NC. NC, negative control; TTN-AS1, titin-antisense RNA1; miR, microRNA; shRNA, short hairpin RNA; MMP, matrix metalloproteinase.

Fig 5: Cordycepin and cisplatin synergistically inhibit osteosarcoma cell invasion by downregulating MMP-2 and MMP-9 expression. a Transwell assay showed that the invasion abilities of U2OS and 143B cells were inhibited following treatment with cordycepin (100 µM), cisplatin (0.5 µM), or their combination for 48 h. Representative images and quantification of the transwell results are shown; b Western blotting analysis showing the protein expression levels of MMP-2 and MMP-9 in U2OS and 143B cells treated with vehicle (Control), cordycepin (200 µM), cisplatin (1 µM), or their combination for 48 h. The data are expressed as the mean ± SD of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001, by Student’s t-test, scale bars = 100 μm