Figure 2.
Modest knockdown of BAFF-R expression sensitizes MCL cells to cytarabine treatment in vitro. (a) Jeko-1 cells were infected with lentivirus shGFP, shBR3-3, or shBR3-4 for 72 hours. The transcriptional level of BAFF-R in Jeko-1 cells expressing shGFP, shBR3-3, or shBR3-4 was determined by RT-qPCR. ***p < .001 for Jeko-1 cells expressing shGFP compared to shBR3-3 and shBR3-3 compared to shBR3-4. (b) Graphical representation (on the left panel) showing the total population of early (Annexin V) and late apoptotic cells (PI) in one of three experiments with triplicate samples 96 hours after shGFP, shBR3-3, or shBR3-4 lentivirus infection in Jeko-1. The quantification data was shown on the right panel. ***p < .001 for Jeko-1 cells expressing shBR3-4 compared to shGFP; ns for Jeko-1 cells expressing shBR3-3 compared to shGFP. (c, left) 1 × 105 Jeko-1 cells or Mino cells (d, left) expressing shGFP or shBR3-3 were seeded in 24-well plate in triplicates and the cell numbers were plotted. p > .05 is considered not significant (ns). Cell counts from (c, right) Jeko-1 shGFP and shBR3-3 cells or (d, right) Mino cells shGFP and shBR3-3 treated with cytarabine ***p < .001. One of the three experiments done with triplicate samples shown. (e, left) The apoptotic cell population was stained by Annexin V and PI followed by flow cytometry analysis Dot plot graph represents the total population of early and late apoptotic cells from (c). Apoptotic cell quantification data shown on the right panel. ***p < .001 for cells expressing shGFP and CYT treatment for 96 hours compared to cells expressing shGFP alone. ***p < .001 for cells expressing shBR3-3 compared to cells expressing shGFP after CYT treatment for 96 hours. Error bars are from single experiment done with triplicate samples representative of three such experiments done with triplicates. (f) Western blot analysis for the apoptotic marker of cleaved caspase-3 in Jeko-1 cells expressing shGFP or shBR3-3 after CYT treatment for 96 hours. GAPDH was used as a loading control