Total cellular lysate was denaturated in 10% SDS and subsequently subjected to IP of ERKI

Cell lysates generated as for quantitative immunoblotting were diluted 1:16 in Array Buffer Plus (Array Buffer composed by 1% BSA, 0,5% NP40, 0,02% SDS, 50mM Tris pH7,4, 150mM NaCl, 1mM EDTA, 5mM NaF, 1mM Na3VO4) containing complete protease-inhibi-tor cocktail (Roche), supplemented with SDS and denatured.

Slides were incubated with calibrators and cell lysates shaking at 4°C overnight.

Cell lysates were diluted 1:10 with Array Buffer PLUS.

B) Total cellular lysate was used to detect phosphorylation of MEK1/2, ERK1/2 and Akt on serine 473 and total actin used as normalizer.

C) Total cellular lysate was used to detect p9ORSK phosphorylation on serine 227 and 380, total p9ORSK and protein disulfide-isomerase (PDI) used as normalizer.

Total cellular lysate was used to detect MEK1/2 phosphorylation, total ERK, total MEK1/2 and total actin used as normalizer.

In other words, a low density population of moderate ESBL producers would lyse entirely because its collective ESBL concentration would be insufficient to inactivate the B-lactam, while a high density population would only eXperience partial lysis before its collective ESBL concentration can inactivate the B-lactam and promote the recovery of the surviving bacteria.

Shortly after the antibiotic is first applied, the population will be reduced due to lysis and appear susceptible because it will not have yet benefited from the activity of ESBLs.

Ideally, a treatment could pinpoint the time window when the most lysis has occurred and the least benefit has been experienced.

In this model, introduction of the antibiotic inhibits bacterial growth and causes lysis .

With a dense enough population, the collective intracellular and extracellular Bla, due to lysis or leaky secretion [50], will be sufficient to degrade the antibiotic to a sublethal concentration before all cells are eliminated (Fig.

If examined shortly after antibiotic treatment, the population will have just experienced significant lysis and will appear susceptible because the effects of Bla have not yet been fully recognized.

Once the initial antibiotic concentration is high enough to cause cell lysis (do > 0.5), the recovery time increases eXponentially with the initial antibiotic concentration until the antibiotic concentration is too high (610 > 10) and the recovery time becomes infinite.

This dependence is an intrinsic property of antibiotic-mediated lysis .

Under increasing concentrations of antibiotic (do > 10), the rate of antibiotic degradation is essentially saturated (limited by the population size and the constant production rate of Bla) and the recovery time is primarily determined by the lysis rate.

Embeddings were first performed on data from cell lysates from 21 neuroblastoma cell lines grown in culture without treatments.

Phosphoproteomic analysis was performed on two endosome (E1, E2) and lysosome ( Lys ) fractions as shown in 810 Fig mass-density plots.

Increased FYN and LYN in faster sedimenting organelles ( lys and E1 fractions) likely reflects migration to multivesicular bodies, late endosomes, and possibly lysosomes [32].

These data were used to define fractions shown on the graph at right ( lys , lysosomes; E1 and E2, endosomes).