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. 2014 Jun 1;20(3):250–257. doi: 10.1089/mdr.2014.0082

FIG. 5.

FIG. 5.

The fusion protein GFP-PBP2xA707D is a target of HtrA. The protein GFP-PBP2xA707D (DKL22) was analyzed in presence or absence of HtrA. (A) Western blot analysis using anti-GFP (α-GFP) antibodies. The positions of GFP-2x* (GFP-PBP2x and GFP-PBP2xA707D) are indicated on the right. Cells were grown in the presence or absence of ZnCl2 (indicated + and −). Arrows point to degradation products that are barely visible in case of ΔhtrA. (B) Western blot of cell lysates developed with anti-PBP2x antibodies. + and −: growth with and without ZnCl2. The position of GFP-2x* and PBP2x are marked. Arrow: product seen in ΔhtrA background. (C) PBP profile analysis. PBPs in cell lysates were labeled with Bocillin FL and visualized by fluorography after separation by SDS-PAGE. On the right the positions of PBPs are indicated. Strains as indicated on top were grown in the presence (+) or absence (−) of ZnCl2. (D) Localization of GFP-PBP2xA707D in the absence of HtrA. DKL22ΔhtrA was grown in the presence of ZnCl2. Left: Phase contrast microscopy; Right: Fluorescence signal. Scale bar=2 μm.