rabbit polyclonal antibody snail1  (Cell Signaling Technology Inc)

Journal: Scientific Reports

Article Title: PAFAH1B3 is a KLF9 target gene that promotes proliferation and metastasis in pancreatic cancer

doi: 10.1038/s41598-024-59427-3

Figure Lengend Snippet: PAFAH1B3 affects the expression of EMT-related proteins in pancreatic cancer cells ( A ) SW1990 and MIA Paca-2 cells were transduced with lentivirus containing PAFAH1B3-Flag or NC, and the protein expression levels of PAFAH1B3, E-cadherin, N-cadherin, Vimentin, Snail1 and MMP2 were measured via Western blotting. ( B ) SW1990 and MIA Paca-2 cells were transduced with lentivirus containing sh-PAFAH1B3 or sh-NC, and the protein expression levels of PAFAH1B3, E-cadherin, N-cadherin, Vimentin, Snail1 and MMP2 were measured via Western blotting. β-Actin was used as an internal control. The data represent the average of three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The following antibodies were used: rabbit anti-PAFAH1B3 polyclonal antibody (1:1000; Abcam, Cambridge, MA, USA), mouse anti-β-actin polyclonal antibody (1:1000; Boster Biological Technology, Ltd), rabbit anti-PCNA polyclonal antibody (1:1000; Abcam, Cambridge, MA, USA), rabbit anti-E-cadherin polyclonal antibody (1:1000; Boster Biological Technology, Ltd), rabbit anti-N-cadherin polyclonal antibody (1:1000; Boster Biological Technology, Ltd), rabbit anti-Snail1 polyclonal antibody (1:1000; Abcam, Cambridge, MA, USA), rabbit anti-MMP2 polyclonal antibody (1:1000; Abcam, Cambridge, MA, USA), rabbit anti-KLF9 polyclonal antibody (1:1000; Abcam, Cambridge, MA, USA), and mouse anti-vimentin polyclonal antibody (1:1000; Boster Biological Technology, Ltd.).

Techniques: Expressing, Transduction, Western Blot

Journal: Bone

Article Title: FBXO11 regulates bone development

doi: 10.1016/j.bone.2023.116709

Figure Lengend Snippet: FBXO11cKO in osteoblastic cells caused osteogenic inhibition through Snail accumulation (6A) shows endogenous Snail1 protein level is lower in FBXO11 overexpression MC3T3 cells, while (6B) shows Snail1 protein level is higher in FBXO11-knockdown cells based on the change in relative protein level observed using western blotting. FBXO11-overexpressing MC3T3 cells and controls were pretreated with MG132 or vehicle for 6 h, and proteins were extracted, IP was performed with anti-Snail1 antibody, and immunoblotting with anti-ubiquitin antibody. Snail1 protein was degraded by FBXO11 through ubiquitination (6C). Snail1 overexpression in osteoblasts snail1 inhibits Overexpression their of osteogenic Snail1 in differentiation. MC3T3 was confirmed by WB with anti-Snail1 antibody (6D). Snail1 overexpression MC3T3 and its control were cultured in osteogenic induction medium in vitro. ALP staining was performed in 0, 7 day group, and AR mineralization staining was performed in 0, 14 day groups. mRNA expression of osteogenic marker genes Runx-2, OSX, Alp, and BSP were compared between Snail1 overexpression MC3T3 cells and their controls in 0, 3 days osteogenic induction groups. The weaker osteogenic differentiation was exhibited in in Snail1 overexpressing osteoblasts. (6E, 6F). In mouse femur bone, the osteoblasts from FBXO11cKO mice exhibits stronger Snail1 immunostaining that WT controls (6G). (X40). In Fig. 5F, all the experiments were repeated three times, the data present as Mean + SD, T-Test; *, P < 0.05; **, P < 0.01. V = control, shFBXO = FBXO11-shRNA.

Article Snippet: We obtained the primary antibodies from the following sources: Rabbit polyclonal antibody to FBXO11 (Novus Biologics, Centennial, CO), rabbit polyclonal antibody to Snail1 (Cell Signaling, Danvers, MA), mouse monoclonal antibody to α-tubulin, and mouse monoclonal antibody to GAPDH (Sigma-Aldrich, St. Louis, MO).

Techniques: Inhibition, Over Expression, Knockdown, Western Blot, Control, Cell Culture, In Vitro, Staining, Expressing, Marker, Immunostaining, shRNA