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. 2022 Jul 29;13:4395. doi: 10.1038/s41467-022-32127-0

Fig. 3. GBP1 peptide binds to and kills F. novicida.

Fig. 3

a Analysis of AMP probability (red) and charge (black) for the mouse GBP1 protein sequence and illustration of the location of putative antimicrobial stretches within the predicted mouse GBP1 structure. b Viability of F. novicida [F. nov., as percentage of CFU in relation to solvent control (Sol.Ctrl.)] assessed 6 h after incubation with GBP128–67, GBP1209–238, GBP1424–452, GBP1558–577 or WLBU2 at 0.1, 1 or 10 μg/mL. c Viability of F. nov. following incubation with GBP128–67 for 6 h over a concentration range (0.01–20 µg/mL) to determine the half maximal inhibitory concentration (IC50). d Flow cytometric analysis (left) and quantitation of flow cytometry plots (right) of SYTOX stained F. novicida treated with Sol.Ctrl. or 100 μg/mL of GBP128–67, GBP1209–238, GBP1424–452, GBP1558–577 or WLBU2 for 12 h. e Confocal microscopy analysis of Hoechst-stained total bacteria (blue), FITC-GBP128–67 (green) and SYTOX (red) in F. novicida or E. coli treated with 10 µg/mL FITC-GBP128–67 or FITC-control peptide for 6 h. White arrows indicate dead bacteria covered with FITC-GBP128–67. f Flow cytometric quantitation of SYTOX stained F. novicida treated with 10 µg/mL of FITC-GBP128–67 for 6 h. g Quantitation of FITC-GBP128–67 bound to F. novicida and E. coli after 1 h incubation with 10 µg/mL of either FITC- GBP128–67 or a FITC-control peptide (in relative fluorescence units, RFU). h Quantitation of the effect of 1 M NaCl, 0.01% saponin or 0.08% sarcosyl on FITC-GBP128–67 binding to F. novicida. Scale bar, 5 µm (e). ns no statistical significance; *P < 0.05, **P < 0.01; ***P < 0.001, ****P < 0.0001 (one-way ANOVA with Dunnett’s multiple-comparisons test (b, d, h), two-tailed t-test (f, g). Data are representative of three independent experiments (b–h; mean and s.e.m. in b–d, f–h). Source data are provided as a Source data file.