Fig. 9
Sorcin mediated the nuclear translocation of PAX5 to regulate FBXL12 transcription. (A) GO analysis of proteins whose expression significantly changed after sorcin was knocked down by shRNA according to the proteomics results. (B) GO proteomics analysis of proteins whose expression significantly changed after sorcin overexpression. (C) Schematic illustration of the cloned fragments in the human FBXL12 promoter. (D) Luciferase activity of the FBXL12 promoter fragments in PANC-1 and Capan-2 cells. (E) Base frequency rule and possible transcription factor-binding site analysis. (F) Ch-IP analysis of the five predicted PAX5 binding sites in the FBXL12 promoter sequence. IgG was used as a negative control. (G) Luciferase activity of the P6 promoter after mutation of site 2 or site 4 in PANC-1 and Capan-2 cells. (H) Western blotting analysis of PAX5, FBXL12 and ALDH1A1 expression in PANC-1, Capan-2 and AsPC-1 cells after PAX5 knockdown or overexpression. (I) Analysis of the interaction between PAX5 and sorcin after transfection with the pCMV-Flag-sorcin vector by coimmunoprecipitation with an anti-Flag antibody followed by Western blotting analysis. (J) Analysis of the interaction between PAX5 and sorcin after transfection with the pCMV-Flag-PAX5 vector by coimmunoprecipitation with an anti-Flag antibody followed by Western blotting analysis. (K) Immunofluorescence analysis of PAX5 nuclear translocation in sorcin-knockdown xenograft tumor tissues. (L) Statistical analysis of the immunofluorescence intensity of PAX5 nuclear translocation in sorcin-knockdown xenograft tumor tissues. (M) Western blotting analysis of PAX5 expression in the nucleus and cytoplasm of PANC-1, AsPC-1 and Capan-2 cells after sorcin knockdown and overexpression. (N) Immunofluorescence analysis of PAX5 levels in the nucleus and cytoplasm in PANC-1 cells after sorcin knockdown and overexpression