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. Author manuscript; available in PMC: 2013 Jan 13.

Published in final edited form as: Science. 2012 Jun 7;337(6091):195–199. doi: 10.1126/science.1219723

Fig. 3 — Human MMS19 interacts with and assembles Fe/S proteins involved in DNA metabolism. HeLa cells were RNAi-treated thrice at three day intervals or mock-treated. Cell extracts were prepared by digitonin lysis (cf. figs. S5–S10). (A) 9 days after starting MMS19 depletion, IRP1 (cytosolic aconitase) activities (normalized to lactate dehydrogenase; LDH) were measured and IRP1, GPAT and actin protein levels were determined by immunostaining. (B) DPYD activity was measured using thin layer chromatography and autoradiography. As controls, HeLa cells were RNAi-depleted for frataxin (FXN), NFS1, and NBP35. For all depletions p<0.001. (C) DPYD and (D) POLD1 protein levels were measured by immunostaining in cell extracts depleted for the indicated proteins. (E) Interaction of human MMS19 with Fe/S proteins and CIA components. Flp-In^™TREx^™-293 cells lacking or stably expressing inducible 3xHA-3xFLAG-MMS19 were induced with doxycycline (500 ng/mL) overnight. Whole cell extracts (WCE) were subjected to anti-HA immunoprecipitation (IP:HA) followed by immunostaining for indicated proteins.

Fig. 3 — Human MMS19 interacts with and assembles Fe/S proteins involved in DNA metabolism. HeLa cells were RNAi-treated thrice at three day intervals or mock-treated. Cell extracts were prepared by digitonin lysis (cf. figs. S5–S10). (A) 9 days after starting MMS19 depletion, IRP1 (cytosolic aconitase) activities (normalized to lactate dehydrogenase; LDH) were measured and IRP1, GPAT and actin protein levels were determined by immunostaining. (B) DPYD activity was measured using thin layer chromatography and autoradiography. As controls, HeLa cells were RNAi-depleted for frataxin (FXN), NFS1, and NBP35. For all depletions p<0.001. (C) DPYD and (D) POLD1 protein levels were measured by immunostaining in cell extracts depleted for the indicated proteins. (E) Interaction of human MMS19 with Fe/S proteins and CIA components. Flp-In^™TREx^™-293 cells lacking or stably expressing inducible 3xHA-3xFLAG-MMS19 were induced with doxycycline (500 ng/mL) overnight. Whole cell extracts (WCE) were subjected to anti-HA immunoprecipitation (IP:HA) followed by immunostaining for indicated proteins.