Anti-TIMP2 Rabbit Monoclonal Antibody

Western blot analysis of TIMP2 expression in HeLa cell lysate.

Effect of LASS2 on cell migration, invasion and apoptosis. (A) Wound healing assay of pLV-vector or pLV-LASS2-transfected glioma/glioblastoma cells at 0 h, 12 h and 24 h after scratch. The images were taken from an inverted microscope under 10× magnification (*P < 0.05 and **P < 0.01, vs. pLV control; unpaired two-tailed Student's t-test; n = 3). Scale bar = 200 µm. (B) Colony formation assay in pLV-vector or pLV-LASS2-transfected U251 and U-87 MG cells. Images were acquired at 4× magnification (*P <0.05, vs. pLV control; unpaired two-tailed Student's t-test; n = 3). (C) Transwell assay demonstrated that LASS2 inhibits the migration of U251 and U-87 MG cells compared with the pLV control (*P <0.05; unpaired two-tailed Student's t-test; n = 3). Scale bar = 200 µm. (D) The immunofluorescence staining of MMP9 and SPHK1 was shown in both U251 and U-87 MG cells. Scale bar = 200 µm. (E) RNA-Seq shows that LASS2 influenced cell migration/invasion, apoptosis, epithelial- mesenchymal transition (EMT) conversion and cellular life activity. (F) Overexpression of LASS2 reduced the protein levels of MMP2, MMP9, and SPHK1 while increasing that of TIMP2 in both U251 and U-87 MG cells. (G) LASS2 overexpression increased the levels of Bax, cleaved Caspase-3, TNF-α, and p53 while reducing that of Bcl-2 in both U251 and U-87 MG cells. (H) Overexpression of LASS2 reduced the protein levels of Vimentin and N-cadherin while increasing that of E-cadherin in both U251 and U-87 MG cells (F, G, and H, *P <0.05 and **P < 0.01, vs. pLV control in each cell line; unpaired two-tailed Student's t-test; n = 3).
Index in PubMed under a CC BY license. PMID: 35517425

LASS2 inhibited tumor growth in a pLV-LASS2-U-87 MG glioblastoma xenograft nude mouse model. (A) Representative photographs showing the gross pLV-LASS2-U-87 MG and empty scramble control glioblastoma xenografts from the nude mouse. (B) The tumor volume was evaluated between the scrambled control and pLV-LASS2 groups (**P < 0.05 and **P < 0.01 vs. pLV control group; unpaired two-tailed Student's t-test; n = 5 animals). (C) The final tumor weight was measured after dissection. The average final weight of tumors derived from pLV-LASS2-transcfected U-87 MG cells was significantly lower than those derived from the scrambled control (**P < 0.01, vs. pLV control group; unpaired two-tailed Student's t-test; n = 5 animals). (D) Representative images for H&E staining from either group were shown. (E) IHC staining of LASS2, TIMP2, MMP9, and SPHK1 in xenografted tumors derived from U-87 MG cells transfected with either pLV-LASS2 or scrambled control. Scale bar = 20 µm. (F) Western blot analysis of LASS2, SPHK1, TIMP2, MMP2, and MMP9 in xenografted tumors derived from U-87 MG cells transfected with either pLV or pLV-LASS2. (G) Western blot analysis of Bax, Bcl-2, pro-Caspase-3, cleaved Caspase-3, TNF-α and p53 in xenografted tumors derived from U-87 MG cells transfected with either pLV or pLV-LASS2. (H) Western blot analysis of EMT conversion-related proteins Vimentin, E-cadherin, and N-cadherin in xenografted tumors derived from U-87 MG cells transfected with either pLV or pLV-LASS2 ( F, G, and H, *P <0.05, **P < 0.01, and ***P < 0.001, vs. pLV control; unpaired two-tailed Student's t-test; n = 3).
Index in PubMed under a CC BY license. PMID: 35517425