Figure 2
From: The α-helical regions of KERP1 are important in Entamoeba histolytica adherence to human cells
KCS is expressed in E. histolytica transfectants and localizes in the same cellular compartments as KERP1.
Biochemical detection of KERP1 and KCS in transfected trophozoites TetOCat and TetOCat-KCS. (a). Immunoblotting of parasite protein extracts at 48 hours of tetracycline induction (+) or without (−). Detection of endogenous actin (49 kDa; mAb anti-actin C4) was used as protein extract loading control. Detection of KERP1 (25 kDa; mAb anti-KERP1 C2-7) in all strains was observed, but detection of HSV-KCS (15 kDa; mAb anti-HSVtag) was only remarked in TetOCat-KCS (+). Immunoblotting was performed using simultaneously all the primary antibodies. (b). Endogenous KERP1 colocalizes with KCS in live parasites. Cellular detection of KERP1 and KCS in transfected trophozoites TetOCat and TetOCat-KCS after 48 hours of tetracycline induction. Micrographs showing immunofluorescence images acquired by confocal microscopy. KERP1 is detected by a specific monoclonal antibody (green; mAb C2-7), KCS by the HSV-tag (red; mAb anti-HSVtag) and nuclei are labeled with DAPI (blue). KERP1 and KCS share the same cellular compartment as observed by the concentrated patches (yellow) at the trophozoite plasma membrane and internal vesicles. Focal planes from a Z-stack were selected. Scale bar 10 µm.
