Supplementary Data
1
Nanoscale analysis of the effects of antibiotics and CX1 on Pseudomonas aeruginosa
2
multidrug-resistant
3
C. Formosa
4
R. E. Duval
5
1
6
(LAAS), Toulouse, France
7
2
Centre National de la Recherche Scientifique, Toulouse, France
8
3
Université de Toulouse, Toulouse, France
9
4
SRSMC, Université de Lorraine - CNRS, Nancy, France
10
5
ABC Platform , Nancy, France
11
6
Laboratoire de Bactériologie Hygiène, Institut Fédératif de Biologie, Toulouse, France
1,2,3,4
4,5
6
, M. Grare , E. Jauvert
and E. Dague
1,2,3*
1,2,3
, A. Coutable
1,2,3
4
, J. B. Regnouf-de-Vains , M. Mourer
4
.
Centre National de la Recherche Scientifique, Laboratoire d’Analyse et d’Architecture des systèmes
®
12 13
* Corresponding author: Etienne Dague, LAAS-CNRS, 31000 Toulouse, France, [email protected], phone:
14
0033561337841
15 16
1
Supplementary Data 1 Antibiotic susceptibility profile of PaR3 (obtained with automated Vitek2 system, BioMérieux, France).
Antibiotic MIC
Results
Ticarcillin
≥ 128
Resistant
Ticarcillin + clavulanic acid
≥ 128
Resistant
Piperacillin
≥ 128
Resistant
Piper + tazobactam
≥ 128
Resistant
Ceftazidim
4
Sensitive
Aztreonam
16
Sensitive
Imipenem
≥ 16
Resistant
Meropenem
≥ 16
Resistant
Tobramycin
≥ 16
Resistant
Gentamicin
≥ 16
Resistant
Amikacin
4
Sensitive
Minocyclin
4
Resistant
Colistin
≤ 0,5
Sensitive
Trimetoprim – Sulfamet.
≥ 320
Resistant
Pefloxacin
4
Intermediate
Ciprofloxacin
1
Sensible
Cefepim
16
Intermediate
Supplementary Data 2
Imaging of P. aeruginosa ATCC 27853 and PaR3. Height images (z-range = 1 µm) recorded on a, native P. aeruginosa ATCC 27853 cells, e, ticarcillin-treated cells (4 µg/mL), i, tobramycin-treated cells (0.25 µg/mL) and m, Cx1-treated cells (16 µg/mL). b, f, j, and n, vertical deflection images corresponding to the previous height images. Height images (zrange = 1 µm) of c, native PaR3 cells, g, ticarcillin-treated cells (4 µg/mL), k, tobramycintreated cells (0.25 µg/mL) and o, Cx1-treated cells (32 µg/mL). d, h, l, p, vertical deflection images corresponding to the previous height images. Vertical cross-sections taken along the solid lines on native cells (black lines), ticarcillin-treated cells (red lines), tobramycin-treated cells (blue lines) and Cx1-treated cells (green lines) on q, P. aeruginosa ATCC 27853, and s, PaR3. Horizontal cross-sections taken along the dashed lines on r, P. aeruginosa ATCC 27853 and t, PaR3.
Supplementary Data 3
Imaging of P. aeruginosa ATCC 27853 and PaR3. a, schematic representation of the PSD analysis. b, Root Mean square formula. Height images (z-range = 15 nm) recorded on c, P. aeruginosa ATCC 27853 native cells, d, ticarcillin-treated cells (4 µg/mL), e, tobramycintreated cells (0.25 µg/mL), and f, Cx1-treated cells (16 µg/mL). Height images (z-range = 15 nm) recorded on g, PaR3 native cells, h, ticarcillin-treated cells (4 µg/mL), i, tobramycintreated cells (0.25 µg/mL), and j, Cx1-treated cells (32 µg/mL). Panel k shows the roughness measured on the height images of P. aeruginosa ATCC 27853 native cells (black line), ticarcillin-treated cells (red line), tobramycin-treated cells (blue line) and Cx1-treated cells (green line). Panel l shows the roughness measured on the height images of PaR3.
Supplementary Data 4
Mapping of P. aeruginosa cell surface elasticity. a, vertical deflection image of native cells, e, ticarcillin-treated cells (4 µg/mL), i, tobramycin-treated cells (0.25 µg/mL). m, height image (z-range = 800 nm) of Cx1-treated cells (32 µg/mL). b, f, j and n, elasticity maps (zrange = 1.5 MPa) corresponding to the vertical deflection images. c, g, k and o, local elasticity maps (z-range = 800 kPa) recorded on one bacterium from the corresponding vertical deflection images. d, h, l and p, distributions of Young Modulus values corresponding to the local elasticity maps.
Supplementary Data 5
POPE:POPG (2:1) supported bilayers. a, height images (z-range = 1.5 nm) of POPE:POPG supported bilayers at different times (0, 1, 2 and 3h) withtout any treatment. b, height images (z-range = 1.5 nm) of POPE:POPG supported bilayers treated by Cx1 (32 µg/mL) at different times (0, 1, 2 and 3h).
Nanoscale analysis of the effects of antibiotics and CX1 on Pseudomonas aeruginosa
2
multidrug-resistant
3
C. Formosa
4
R. E. Duval
5
1
6
(LAAS), Toulouse, France
7
2
Centre National de la Recherche Scientifique, Toulouse, France
8
3
Université de Toulouse, Toulouse, France
9
4
SRSMC, Université de Lorraine - CNRS, Nancy, France
10
5
ABC Platform , Nancy, France
11
6
Laboratoire de Bactériologie Hygiène, Institut Fédératif de Biologie, Toulouse, France
1,2,3,4
4,5
6
, M. Grare , E. Jauvert
and E. Dague
1,2,3*
1,2,3
, A. Coutable
1,2,3
4
, J. B. Regnouf-de-Vains , M. Mourer
4
.
Centre National de la Recherche Scientifique, Laboratoire d’Analyse et d’Architecture des systèmes
®
12 13
* Corresponding author: Etienne Dague, LAAS-CNRS, 31000 Toulouse, France, [email protected], phone:
14
0033561337841
15 16
1
Supplementary Data 1 Antibiotic susceptibility profile of PaR3 (obtained with automated Vitek2 system, BioMérieux, France).
Antibiotic MIC
Results
Ticarcillin
≥ 128
Resistant
Ticarcillin + clavulanic acid
≥ 128
Resistant
Piperacillin
≥ 128
Resistant
Piper + tazobactam
≥ 128
Resistant
Ceftazidim
4
Sensitive
Aztreonam
16
Sensitive
Imipenem
≥ 16
Resistant
Meropenem
≥ 16
Resistant
Tobramycin
≥ 16
Resistant
Gentamicin
≥ 16
Resistant
Amikacin
4
Sensitive
Minocyclin
4
Resistant
Colistin
≤ 0,5
Sensitive
Trimetoprim – Sulfamet.
≥ 320
Resistant
Pefloxacin
4
Intermediate
Ciprofloxacin
1
Sensible
Cefepim
16
Intermediate
Supplementary Data 2
Imaging of P. aeruginosa ATCC 27853 and PaR3. Height images (z-range = 1 µm) recorded on a, native P. aeruginosa ATCC 27853 cells, e, ticarcillin-treated cells (4 µg/mL), i, tobramycin-treated cells (0.25 µg/mL) and m, Cx1-treated cells (16 µg/mL). b, f, j, and n, vertical deflection images corresponding to the previous height images. Height images (zrange = 1 µm) of c, native PaR3 cells, g, ticarcillin-treated cells (4 µg/mL), k, tobramycintreated cells (0.25 µg/mL) and o, Cx1-treated cells (32 µg/mL). d, h, l, p, vertical deflection images corresponding to the previous height images. Vertical cross-sections taken along the solid lines on native cells (black lines), ticarcillin-treated cells (red lines), tobramycin-treated cells (blue lines) and Cx1-treated cells (green lines) on q, P. aeruginosa ATCC 27853, and s, PaR3. Horizontal cross-sections taken along the dashed lines on r, P. aeruginosa ATCC 27853 and t, PaR3.
Supplementary Data 3
Imaging of P. aeruginosa ATCC 27853 and PaR3. a, schematic representation of the PSD analysis. b, Root Mean square formula. Height images (z-range = 15 nm) recorded on c, P. aeruginosa ATCC 27853 native cells, d, ticarcillin-treated cells (4 µg/mL), e, tobramycintreated cells (0.25 µg/mL), and f, Cx1-treated cells (16 µg/mL). Height images (z-range = 15 nm) recorded on g, PaR3 native cells, h, ticarcillin-treated cells (4 µg/mL), i, tobramycintreated cells (0.25 µg/mL), and j, Cx1-treated cells (32 µg/mL). Panel k shows the roughness measured on the height images of P. aeruginosa ATCC 27853 native cells (black line), ticarcillin-treated cells (red line), tobramycin-treated cells (blue line) and Cx1-treated cells (green line). Panel l shows the roughness measured on the height images of PaR3.
Supplementary Data 4
Mapping of P. aeruginosa cell surface elasticity. a, vertical deflection image of native cells, e, ticarcillin-treated cells (4 µg/mL), i, tobramycin-treated cells (0.25 µg/mL). m, height image (z-range = 800 nm) of Cx1-treated cells (32 µg/mL). b, f, j and n, elasticity maps (zrange = 1.5 MPa) corresponding to the vertical deflection images. c, g, k and o, local elasticity maps (z-range = 800 kPa) recorded on one bacterium from the corresponding vertical deflection images. d, h, l and p, distributions of Young Modulus values corresponding to the local elasticity maps.
Supplementary Data 5
POPE:POPG (2:1) supported bilayers. a, height images (z-range = 1.5 nm) of POPE:POPG supported bilayers at different times (0, 1, 2 and 3h) withtout any treatment. b, height images (z-range = 1.5 nm) of POPE:POPG supported bilayers treated by Cx1 (32 µg/mL) at different times (0, 1, 2 and 3h).
