FIG 5.

Mce4A and Mce4E interact. A clarified WCL generated by passage through a French pressure cell was used for coimmunoprecipitation with anti-HA-conjugated beads, followed by immunoblot analysis. The amount of clarified WCL (Lys) loaded represents one-fifth of the amount of protein used for the immunoprecipitation (IP) sample. (A) The strains used for immunoprecipitation were M. smegmatis Δmce4A expressing either Mce4A-HA_mc or Mce4A_mc without a tag as a control. An HA antibody was used to detect Mce4A-HA and an Mce4E antibody was used to detect Mce4E in the coimmunoprecipitate. (B) The strains used for immunoprecipitation were M. smegmatis Δmce4E expressing either Mce4E-HA_mc or Mce4E_mc without a tag as a control. An Mce4E antibody was used to detect Mce4E-HA and an Mce4A antibody was used to detect Mce4A in the coimmunoprecipitate. The arrow indicates the Mce4A band, to distinguish it from the higher species cross-reacting band. An MspA antibody was used to detect MspA as a negative control. (C) The strains used for immunoprecipitation were M. smegmatis Δmce4A expressing Mce4A-HA_sc or the WT strain expressing an empty vector (EV) with an HA tag. An HA antibody was used to detect Mce4A-HA and an Mce4E antibody was used to detect Mce4E in the coimmunoprecipitate. (D) The strains used for immunoprecipitation were M. smegmatis Δmce4E expressing Mce4E-HA_sc or the WT strain expressing an empty vector (EV) with an HA tag. An HA antibody was used to detect Mce4E-HA and an Mce4A antibody was used to detect Mce4A in the coimmunoprecipitate. The arrow indicates the Mce4A band, to distinguish it from the higher species cross-reacting band. Mutant strains contain empty vectors (EV) as indicated.