Research Paper Volume 15, Issue 12 pp 5381—5398
FBXO28 promotes proliferation, invasion, and metastasis of pancreatic cancer cells through regulation of SMARCC2 ubiquitination
Figure 4. SMARCC2 is a critical target of FBXO28. (A) BxPC-3 cells with stable FBXO28 overexpression, as well as control cells, were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and band staining for observation. Mass spectrometry was used to identify and isolate protein bands. (B) Liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis of five proteins co-precipitated with FBXO28. (C) Mass spectrometry revealed a unique peptide of FBXO28 identified by two-dimensional LC-MS/MS from protein lysates of anti-FBXO28 immunoprecipitated BxPC-3 cells. (D) The interaction between FBXO28 and SMARCC2 was confirmed using a co-immunoprecipitation assay, and western blots were performed on cell lysates. (E) Co-localization analysis of FBXO28 and SMARCC2 in the nucleus by immunofluorescence. (F, G) Typical immunohistochemical (IHC) image showing low expression of SMARCC2 in pancreatic cancer tissue (magnification: ×50, ×200). (H–J) Immune tissue co-expression revealed a negative association between FBXO28 and SMARCC2 expression within pancreatic cancer tissues (magnification: ×50, ×200). ****P < 0.0001.