Comparison between HsNudT16 and its mutants in their ability to process MARylated PARP10CD. (A) Coomassie blue-stained SDS gel of 32P-MARylated PARP10CD (top band) demodification reaction by HsNudT16 and mutants Δ17, F36A, F61S, F36A F61S and H24W displaying comparable amount of protein PARP10CD at every time point. The positions of the molecular weight standards corresponding to 25 and 20 kDa are shown on the left. (B) 32P autoradiograph of the reactions in (A) displaying a decrease in radioactivity as time elapse. (C) Quantification of removal of 32P-radioactive signal from PARP10, n = 3. Please note that the apparent activity of H24W observed is due to the background hydrolysis of the MARylated substrates as shown in the buffer control (Supplementary Fig. S5).