Fig 3. BCC0 knockdown does not affect the localization of IMC32 or IMC43.

A) Diagram depicting the ATc-regulatable transcriptional repression system used for BCC0 knockdown. The endogenous promoter for BCC0 was replaced with a TetO7-SAG4p minimal promoter in a BCC0^2xStrep3xTy strain expressing the Tati transactivator. B) IFA of BCC0^ckD parasites grown -/+ ATc for 24 hours showing that knockdown of BCC0 results in severe morphological defects. Magenta = anti-Ty detecting BCC0^cKD, Green = anti-IMC6. C) Western blot and quantification showing that BCC0^cKD is 93% depleted after 24 hours of ATc treatment. D) Plaque assays show that both plaque number and size are significantly reduced when BCC0 is knocked down. Statistical significance was determined using a two-tailed t-test (****, P < 0.0001). E) IFA of BCC0^ckD parasites grown -/+ ATc for 24 hours showing that knockdown of BCC0 does not affect IMC32 localization. Magenta = anti-Myc detecting IMC32^3xMyc, Green = anti-IMC6. F) IFA of BCC0^ckD parasites grown -/+ ATc for 24 hours showing that knockdown of BCC0 does not affect IMC43 localization. Magenta = anti-HA detecting IMC43^smHA, Green = anti-IMC6. Scale bars = 2 μm.