Figure 6.—

HA-tagged LF1 is present in the cell body and flagella. (A) Restriction map of 7.2-kb genomic plasmid showing the placement of the triple HA tag within the LF1 gene. (B) Immunoblot using an HA antibody shows four HA-tagged lf1 rescued strains (E1, G9, G12, and G12) express the HA-LF1 tagged protein, whereas an lf1 rescued strain transformed with a plasmid lacking the HA tag is not reactive to the HA antibody. (C) Immunoblot using an HA antibody indicates HA-LF1p is present in whole cells, cell bodies, and flagella. Lanes 1, 2, and 3 show protein from whole cell, cell body, and flagella (30 μg). Lanes 4 and 5 show protein from whole cells and cell bodies (2 × 10⁶). Lanes 6, 7, 8, 9, and 10 show protein from flagella isolated from 2 × 10⁶ (1×), 2 × 10⁷ (10×), 1 × 10⁸ (50×), 2 × 10⁸ (100×), and 2 × 10⁹ (1000×) cells. (D) Immunofluorescence of HA-LF1 tagged cells. Shown are phenotypically rescued strains, G2 and E1, stained with anti-HA antibody (1 and 3) and anti-tubulin antibody (2 and 4) and wild-type cells lacking the HA tag stained with anti-HA (5) and anti-tubulin (6) antibodies.