ssGPL does not contribute to the resistance of <i>M</i>. <i>avium</i> to LL-37.

<p>CFU determination of <i>M</i>. <i>avium</i>^ssGPL and <i>M</i>. <i>avium</i>^ΔssGPL serovar 8 after incubation with 0, 10, 25, and 100 μg/ml LL-37 and 20 μg/ml gentamicin. ***p<0.0001. Data are the mean ± SEM of 3 independent experiments.</p

Loss of LL-37 activity after exposure to NTM or NTM-derived lipids.

<p><i>E</i>. <i>coli bioassays</i> were used to evaluate LL-37 activity. (A) LL-37 remaining in NTM (but not <i>Mtb</i>) culture supernatant no longer kills <i>E</i>. <i>coli</i>. (B) <i>E</i>. <i>coli</i> survives in untreated or boiled NTM culture supernatants to which fresh LL-37 was added. (C) <i>E</i>. <i>coli</i> survival following incubation with <i>M</i>. <i>abscessus</i> or <i>M</i>. <i>intracellulare</i> derived cell fractions. CM = cell membrane, CW = cell wall, ICW = insoluble cell wall fraction.</p

<i>E</i>. <i>coli</i> is susceptible to LL-37.

<p>(A) Log₁₀ CFU of <i>E</i>. <i>coli</i> after 4 hours of incubation with 0, 10, or 25 μg/ml of LL-37. **p< 0.001; ***p<0.0001. Data are the mean ± SEM of 6 independent experiments. (B) Images were taken of each serial dilution on LB agar from <i>E</i>. <i>coli</i> cultures incubated for 4 hours in the absence or presence of 25 μg/ml of LL-37.</p

LL-37 demonstrates broad-spectrum antimicrobial activity.

<p>Log₁₀ CFU after 1–8 hours of incubation of a (A) laboratory isolate of <i>Salmonella enteriditis</i> or clinical isolates of (B) <i>Salmonella enteriditis</i> (Uganda) or (C) <i>Salmonella non-typhi</i> (Nairobi) with 0–50 μg/ml of LL-37. *p< 0.01; **p< 0.001; ***p<0.0001. Data are the mean ± SEM of 3–6 independent experiments. (D) <i>Mtb</i> H37Rv were incubated with 10 μg/ml LL-37 and the percent change in CFU calculated after 96 hours incubation. n = 3 independent experiments.</p

nsGPL do not mediate the resistance of <i>M</i>. <i>abscessus</i> to LL-37.

<p>(A-B) CFU determination of <i>M</i>. <i>abscessus</i>^nsGPL(+) in the presence of the indicated concentrations of native LL-37 (p = 0.69). Data are the mean ± SEM of 4 independent experiments. (B) CFU determination of <i>M</i>. <i>abscessus</i>^nsGPL(-) in the presence of the indicated concentrations of native LL-37. Data are the mean ± SEM of 4 independent experiments. *p<0.01; **p< 0.001; ***p<0.0001. (C) Thin-layer chromatography demonstrates the presence and absence of GPL in <i>M</i>. <i>abscessus</i>^nsGPL(+) and <i>M</i>. <i>abscessus</i>^nsGPL(-), respectively. (D-E) CFU determination of <i>M</i>. <i>abscessus</i>^nsGPL(+) and <i>M</i>. <i>abscessus</i>^nsGPL(-), respectively in the presence of the indicated concentrations of scrambled LL-37 peptide. Data are the mean ± SEM of 3 independent experiments. *p<0.01; **p< 0.001; ***p<0.0001.</p