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. 2017 Oct 12;12(10):e0186187. doi: 10.1371/journal.pone.0186187

Fig 5. Western blotting and SDD-AGE analysis of Hrp1 and related chimera.

Fig 5

Western blots were used to examine expression of Hrp1. Whole cell lysates from strains DY388, DY387, DY387F, DY389, DY3242, and DY3243 were boiled in loading buffer, resolved by SDS-PAGE, transferred to nitrocellulose, and probed with anti-Hrp1 antibody, as described in Methods. Pre-stained marker proteins were run in an adjacent lane and their sizes are indicated at left. To the right are SDD-AGE assays analyzing lysates from strains bearing the indicated Hrp1-derivatives (lane 7 = DY308, lane 8 = DY3243, lane 9 = DY4500, lanes 10–12 = DY387F, lanes 13 and 14 = DY388, lane 15 and 16 = DY387F), and developed with anti-Hrp1 antibody, as described in Methods. Lanes 7–12 (all lanes are derived from the same gel and filter) show that the oligomeric forms of Hrp1CT25 (“*”) are specific for that Hrp1 derivative, and that the proteins are relatively resistant to reducing agent. Lanes 13–16 show that the oligomers (“*”) and high molecular weight (MW) forms of Hrp1 and Hrp1CT25 are relatively resistant to boiling in SDS for 5 min before electrophoresis.