Fig. 1.

Poly-SUMOylated proteins accumulate during mechanical ventilation treatment in rat diaphragms. A and C, muscle samples listed in supplemental Table 1 were fractionated by SDS-polyacrylamide gel and probed with antibodies to SUMO1 and SUMO2/3. Representative Western blottings of diaphragm muscle lysates from controls (ctrl) and from ventilated rats from 6 h to 1 day (1d), from 1 to 5 days (5d), from 5 to 10 days without (10d), and in the presence of the drug BGP-15 (10d BGP-15) were fractionated by SDS-polyacrylamide gel, and PVDF membranes were probed with anti-SUMO1 (A) and anti-SUMO2 antibodies (C). B and D, quantification of free and conjugated SUMO1 and SUMO2/3. The intensities of free SUMO and SUMO conjugates identified by the dashed areas in A and B were quantified by densitometry in two independent experiments. The intensity band quantifications for each area were expressed in AU after normalization by the correspondent GAPDH loading control and referred to the control samples described in supplemental Table 1.