Fig 2: The effects of AQP3-silencing on the proliferation, invasion and epithelial-mesenchymal transition of osteosarcoma cells. U2OS cells were trans-fected with sti-AQP3 or sti-NC. (A) The protein and mRNA expression of AQP3 was determined by western blot analysis and RT-qPCR, respectively. (B) Cell proliferation was assessed using a BrdU-ELISA assay. (C) The mRNA expression of PCNA, CDK4, cyclin D1 and p27 was determined by RT-qPCR. (D) Invasion was assessed using a Transwell assay. (E) Protein expression of MMP-2, MMP-9 and TIMP-1 was detected by western blot analysis. (F) The expression of E-cadherin, N-cadherin and Vimentin was determined by western blot analysis. All data are presented as the mean ± standard error of the mean, n=6. ##P<0.01 vs. sti-NC. AQP3, aquaporin 3; si, small interfering RNA; NC, negative control; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; BrdU, bromodeoxyuridine; PCNA, proliferating cell nuclear antigen; CDK4, cyclin-dependent kinase 4; MMP, matrix metalloproteinase; TIMP, TIMP metallopeptidase inhibitor; OD, optical density.

Fig 3: CXTB attenuates cartilage injury by downregulating the lncRNA HOTAIR. (A) The expression of lncRNA HOTAIR. (B-G) HE staining of the articular cartilage tissue (×200) and the OARSI scores rats in different treatment groups. (H-L) The protein and mRNA expression of MMP13 and TIMP1. *P<0.05; #P>0.05. CXTB, cangxitongbi; lncRNA, long non-coding RNA; HOTAIR, Hox transcript antisense intergenic RNA; HE, hematoxylin and eosin; OARSI, Osteoarthritis Research Society International; MMP13, matrix metallopeptidase 13; TIMP1, tissue inhibitor matrix metalloproteinase 1; NC, negative control; AAV, adeno-associated virus.

Fig 4: IL12A regulates expression of angiogenesis-related proteins in GC cells, including TIMP1, IGFBP1 and PAI1. (A, B) Angiogenesis-related proteins of indicated HGC27 cells were detected with proteome profiler human angiogenesis array kit. (C) Relative expressions of TIMP1, PAI1 and IGFBP1 were detected by western blotting in pc-IL12A, pc-EBI3 and pc-Ctrl HGC27 cells. (D) Relative expressions of TIMP1, PAI1 and IGFBP1 were detected by western blotting in sh-IL12A, sh-EBI3 and pYr-ctrl AGS cells. (E) TIMP1, PAI1 and IGFBP1 were detected by real-time quantitative PCR in indicated HGC27 and AGS cells. Data are shown as the mean ± SD; statistical test: Student’st-test and one-way ANOVA with the Dunnettpost hoctest;n = 3; *P < 0.05, **P < 0.01. ns, no statistical difference.

Fig 5: TIMP1 expression level was associated with different clinicopathological parameters. The mRNA expression levels of TIMP1 were downloaded from the TCGA-KIRC database. TIMP1 was upregulated in (A) T stage, (B) lymph node metastasis, (C) distant metastases, (D) TNM stage, (E) G grade, (F) gender, (G) OS status, (H) DFS status. ****p < 0.0001; ***p < 0.001; *p < 0.05. TIMP1, Tissue inhibitor matrix metalloproteinase 1; RCC, clear cell renal cell carcinoma; KIRC, kidney renal clear cell carcinoma; TCGA, The Cancer Genome Atlas; TNM, Tumor-Node-Metastasis; OS, overall survival; DFS, disease-free survival.