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. 2022 Oct 5;96(20):e00549-22. doi: 10.1128/jvi.00549-22

FIG 4.

FIG 4

Assay of expression levels of EIAV viral proteins in cells overexpressing SYNJ2BP or KO HEK293T cells. (A to C) Overexpression of SYNJ2BP can promote the production of EIAV Env but not Gag or Rev. HEK293T cells were cotransfected with plasmids, either pcDNA3.1-Flag-SYNJ2BP in three doses or an empty plasmid and pcDNA3.1-Env (EIAV), or pcDNA3.1-Flag-SYNJ2BP and VR1012-Gag (EIAV) or pcDNA3.1-rev-HA (EIAV). Cell lysates were analyzed by using Western blotting to detect the intensity of the EIAV Env or Gag or Rev protein band at 48 hpt. This experiment was performed three times. (D to F) Knockout of SYNJ2BP reduced production of EIAV Env but not Gag or Rev. (D) Empty plasmid or pcDNA3.1-Flag-SYNJ2BP was cotransfected into WT or ΔSYN HEK293T cells, together with pcDNA3.1-Env (EIAV). (E and F) WT and ΔSYN HEK293T cells were transfected with VR1012-Gag (EIAV) or pcDNA3.1-rev-HA (EIAV). Cell lysates were analyzed as in panels A to C. The results of the densitometry analysis to quantify the ratio of Env, Gag, or Rev to β-actin are shown at the bottom (lane 1 set as 1.0). This experiment was performed three times.