Figure 1.

ARTD1 PARylates proteins, including itself, and is degraded during OC formation via the proteasome pathway. (a) Western blot analysis using PAR antibody of protein PARylation from 2% CMG-treated BMM cultures; bracket indicates areas of profound changes in PARylation. (b) Western blot analysis of the effect of vehicle, olap or velip on 2% CMG-induced protein PARylation for 24 h in BMM cultures using PAR antibody. (c) Western blot analysis (using PAR antibody) of protein PARylation during OC differentiation induced by 2% CMG and 100 ng/ml RANKL (experiment was run in duplicates). (d) Time-course effect of CMG-containing M-CSF on ARTD1 expression (h, hours of CMG treatment). (e) Analysis of ARTD1 during OC differentiation from RAW 264.7 cells. (f) Analysis of ARTD1 expression during OC differentiation (d, days of exposure to CMG and RANKL). (g) Analysis of ARTD1 during OC differentiation using GST-Af1521 macrodomains. On day 3, cultures were treated with olap, and the experiment was terminated on day 4 (OC, 4 d, +olap). (h) Analysis of ARTD1 during OC differentiation by immunoprecipitation using PAR antibody. (i) Effect of the proteasome inhibitor, MG-132 on ARTD1 fate. Cultures were treated with 8 μM MG-132 on day 3 for 3 h prior to harvesting samples. Results are from the same gels, but the lanes were cut and pasted. Incubation with MG-132 for >3 h caused cytotoxicity. Data are representative of at least three independent experiments