Migration of BCWM.1 and CXCR4 knockdown cell lines. (A) Transwell migration assay using BCWM.1 and WM-WSU (WSU) WM cell lines. SDF-1 0 to 100 nM was placed in the lower chambers, and migration was determined after 4 hours. Maximum migration occurred at approximately 30 nM with no further increase in migration beyond 30 nM. (B) CXCR4 knockdown BCWM.1 cell line (CXCR4Kd) was generated using lentivirus infection. Flow cytometry for CXCR4 expression was performed using IgG1 isotype control, CXCR4 Kd cell line, and mock BCWM.1 showing no CXCR4 expression in the Kd cell line. Similarly, Western blotting demonstrates negligible CXCR4 expression in the Kd cell line compared with mock BCWM.1 (C) Transwell migration assay using CXCR4Kd cell line compared with Mock-infected BCWM.1 cell line showing significant migration with SDF-1 30 nM, whereas the CXCR4 Kd cell line showed minimal migration in response to SDF-1.
