Figure 1.
Comparison of the MEC1 and MEC2 cells.
(A) Expression of EBV encoded proteins EBNA-2 and LMP-1 by immunofluorescence; magnification (×100), scale bar 25 µm. Note: the MEC2 cells are larger. (B) Expression of EBNA-2 and LMP-1 by immunoblotting; positive control: CBM1-Ral-STO, negative control: Ramos. 1.5×105 cells were loaded in control lanes and 5×105 were loaded in MEC1 and MEC2 lanes. Note MEC2 expresses higher amount of EBNA-2. (C) Expression of Bright and BARF1 by Q-PCR. (D) FACS analysis of surface markers that are differently expressed in the 2 lines.
Figure 2.
The effect of IL-21 and CD40L exposure on MEC1 and MEC2 cells.
Expression of EBNA-2 and LMP-1 in IL-21 treated cells (A, B). (A) Simultaneous immunofluorescence staining of EBNA-2 (Green) and LMP-1 (Red); magnification (×100), scale bar 25 µm. Note the downregulation of EBNA-2 and upregultion of LMP-1 after IL-21 treatment. (B) Expression of EBNA-2, LMP-1 and Blimp-1 by immunoblotting; positive control: CBM1-Ral-STO, negative control: Ramos. 1.5×105 cells were loaded in the control lanes and 5×105 were loaded in both untreated and IL-21 treated MEC1 and MEC2 lanes. Note low expression of EBNA-2 and high expression of LMP-1 after IL-21 treatment and induction of Blimp-1 after IL-21 treatment. (C) Activity of the W and C promoters that regulate EBNA-2 expression and LMP-1 mRNA expression by Q-PCR. Note the difference in EBNA-2 regulation; the MEC2 cell uses both Wp and Cp while in MEC1 only Wp is active. (D) Expression of EBNA-2 and LMP-1 in cells exposed to CD40L. Simultaneous immunofluorescence staining; for details see (A). Note: EBNA-2 and LMP-1 are downregulated by CD40L in both lines. (E) CD40L induced modulation of surface marker by FACS analysis.