Skip to main content
. 2017 Jul 27;13(7):e1006542. doi: 10.1371/journal.ppat.1006542

Fig 8. IE1 AA451-475 is dispensable for Sp100A interaction and downregulation.

Fig 8

(A-B) Effect of Δ451–475 on protein levels of exogenous Hes1 and Sp100A in 293T cells. 293T cells were co-transfected with 0.25μg pCDH-Hes1 (Hes1) (A) or pCMV-Myc-Sp100A (Myc-Sp100A) (B) along with 2.5μg pEYFP (vector), pEYFP-IE1 (IE1) or pEYFP-IE1(Δ451–475) (Δ451–475). Cells were harvested at 48hpt for IB to examine the protein levels of Hes1 (A) or Sp100A (B). The values listed below the blots indicate the relative protein levels of Hes1 or Sp100A compared to the controls following β-actin normalization. Representative images from 3 independent experiments are shown (upper panel), and relative protein levels are presented as the mean ± SD (lower panel). *, P≤0.05; **, P≤0.01. (C) Interaction of IE1 or Δ451–475 with Sp100A in 293T cells. 293T cells were transfected with 6μg pCMV-Myc-Sp100A (Myc-Sp100A) and 6μg pEYFP (vector), pEYFP-IE1 (IE1) or pEYFP-IE1(Δ451–475) (Δ451–475). Cells were harvested at 48hpt and cell lysates were subjected to Myc-directed IP and subsequent IB for IE1 and Myc. Normal IgG was applied as nonspecific antibody control, and the indicated proteins were examined in the cell lysates. The band corresponding to IgG heavy chain is indicated by an arrow.