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. 2009 May 1;10(7):748–754. doi: 10.1038/embor.2009.86

Figure 3.

Figure 3

Silencing of MAPL reduces total SUMO1 conjugates. (A) HeLa cells were stably transfected with either shRNA vector alone (shVector) or with MAPL-specific shRNA (shMAPL). Total lysates were probed for SUMO1. (B) As in (A), but the extracts were probed with anti-ubiquitin antibodies. (C) HeLa cells were transfected with either a pooled siRNA mixture directed against MAPL or a control siRNA. Total lysates were probed for SUMO1. (A–C) The graphs represent the average of three independent experiments. (D) Isolation of the nuclear (Nuc), cytosolic (Cyto) and mitochondrial (Mito) fractions of HeLa cells stably transfected with an shRNA directed against MAPL or a control shRNA. A 40% reduction in the expression of MAPL leads to a 30% reduction in total SUMO1 conjugates, 38% reduction in nuclear SUMO1 conjugates, 20% reduction in cytosolic SUMO1 conjugates and a 53% reduction in mitochondrial SUMO1 conjugates. HSP, heat-shock protein; MAPL, mitochondrial-anchored protein ligase; NT, non-targeted; shRNA, short hairpin RNA; siRNA, short interfering RNA; SUMO, small ubiquitin-like modifier; TOM, translocase of the mitochondrial outer membrane; Tot, total.