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. 2008 Sep 22;155(8):1235–1249. doi: 10.1038/bjp.2008.370

Figure 4.

Figure 4

Effects of mono-galloyl glucose on poly(ADP-ribose) glycohydrolase (PARG) activity in intact cells. (a) HeLa cells were exposed to the poly(ADP-ribose) polymerase (PARP-1)-activating compound 1-methyl-3-nitro-1-nitrosoguanidine (MNNG) (100 μM) in the absence or presence of 10 μM 3-galloyl-glucose (5) and poly(ADP-ribose) (PAR) levels were evaluated by western blotting at different times. β-Actin is shown as a loading control. (b) HeLa cells were exposed to the PARP-1-activating compound MNNG (100 μM) in the absence or presence of 3-galloyl-1,2-O-isopropylidene glucose (4) (10 or 100 μM) and PAR levels were evaluated by western blotting at different time points. β-Actin is shown as a loading control. Compound (4) was pre-incubated for 30 min. (c) Immunocytochemical evaluation of intracellular PAR contents in cells exposed to MNNG in the absence or presence of 3-galloyl-1,2-O-isopropylidene glucose (4, 100 μM). (d) Nicotinimade adenine dinucleotide (NAD) levels in cells exposed 30 min to 30 μM 6(5H)-phenanthridinone (PHE) or 30 and 60 min to 100 μM 3-galloyl-1,2-O-isopropylidene glucose (4). (e) NAD contents in cells exposed to 100 μM MNNG in the presence or absence of PHE (30 μM) or 3-galloyl-1,2-O-isopropylidene glucose (4, 100 μM). A blot/experiment representative of two (a–c) or three (d) is shown. Bar=20 μm. Columns are the mean±s.e.mean of two experiments (e). *P<0.05, **P<0.01 vs control (C) (ANOVA+Tukey's post hoc test).