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. Author manuscript; available in PMC: 2018 Jun 17.
Published in final edited form as: Biochem Biophys Res Commun. 2017 May 5;488(1):153–158. doi: 10.1016/j.bbrc.2017.04.158

Figure 2. Dfi1 is an integral membrane protein when at least one hydrophobic region is present.

Figure 2

Cell lysates of C. albicans strains encoding HA-tagged full-length Dfi1, Dfi1ΔN, Dfi1ΔC, or Dfi1ΔNΔC underwent ultracentrifugation to pellet cell membranes at 300,000g for 20 min, as described in the Materials and Methods. The pellets were then homogenized with extraction buffer alone, buffer plus 4M urea (final concentration), or buffer plus 5% Triton X-100 (final concentration; v/v). These homogenates then underwent a second round of ultracentrifugation and the resulting supernatants (S) and pellets (P) were probed for HA by Western blotting. The lanes from the DF1ΔC strain were each loaded with 12.5 μL of sample, to avoid excessive signal, and all other lanes were each loaded with 50 μL of sample. Each blot is representative of at least three experiments.