Su 1

Justin Su
Doug Bradley
WRIT 159A
Assignment #8
Figure 5 of Ma et al.
This figure represents a blot of protein levels
containing two different biochemical techniques:
whole cell lysate and immunoprecipitate (IP). Whole
cell lysate contains all the protein content in all the
cells that are lysed. IP, however, is a fractional
protein amount of the whole cell lysate that has been
selected by specific antibodies or magnetic beads. In
this experiment, stable HeLa cell lines – WDR5-
EGFP and Green Fluorescent Protein (GFP) as a
control – were lysed, and a part of their whole cell
lysates were fractioned out for IP. During IP, GFP
beads were used to probe the sample for proteins and
protein complexes tagged with GFP, ultimately used
to deduce protein-protein interactions. Both the
whole cell and IP lysates were running through
sodium dodecyl sulfate polyacrylamide gel
electrophoresis (SDS-PAGE), and transferred onto a
nitrocellulose membrane for Western analysis.

As the blots contain separated proteins and protein

complexed via size, shape, and charges, specific
proteins may be probed, and their protein expression
levels may be analyzed. Firstly, GFP was probed
among different lanes containing whole cell lysates
and IP lysates that were fractioned 30 times the
volume of the whole cell lysates. The data not only
show that WDR5 is indeed stably expressed with an
EGFP tag, but also reveal that there is an increase in
WDR5-EGFP expression under infectious
conditions. In addition, RbBP5 is known to associate
with WDR5, yet does not show a strong impact to the
known association upon measles virus (MeV)
infection. Furthermore, MeV L, N, P, and C are
measles virus proteins, and are found in the IP
samples when probed. Having no bands shown in
uninfected conditions, this indicates that WDR5-
EGFP associates with these proteins, either directly
or indirectly. MeV H and V proteins were not found
to be associated with WDR5-EGFP.

This figure is strong evidence that WDR5-EGFP is

significant in the role of measles virus replication.