You are studying $4$ proteins that based on bioinformatics analysis are predicted to function as amylases in a protozoan parasite that causes a sexually transmitted disease. Amylases, are enzymes that can break down glycogen. You clone these proteins into an overexpression plasmid to create HA$-$tagged fusion proteins $($as shown in the schematic$).$ HA is a human influenza hemagglutinin protein that we often use as a molecular tag to detect recombinant proteins.

You treat half of the cells with a vehicle control $(-$ condition$)$ or with PNGase F $(+$ condition$).$ PNGase F is an enzyme that cleaves off Nlinked oligosaccharides from glycoproteins. You spin down the cells at $3,2000$ rpm to pellet the cells. You transfer the liquid part$=$cell supernatant into a separate test tube. You then lyse the remaining cell pellet with detergent to prepare whole cell lysates. You prepare the whole cells lysates and cell supernatants for SDS$-$PAGE and perform western blots using an antibody that recognizes the HA tag on the fusion proteins.

If you need review on western blot click on the "Hint" button.

$$II$-$

Choose one or more:

A$.$ BA$-1$ is phosphorylated

B$.$ BA$-4$ is phosphorylated

C$.$ BA$-1$ is glycosylated

D$.$ BA$-4$ is glycosylated

E$.$ BA$-1$ is secreted

F$.$ BA$-4$ is secreted