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. 2018 May 21;293(28):10937–10948. doi: 10.1074/jbc.RA117.000598

Figure 3.

Figure 3.

Methylated C-terminal end of TLS still binds to its N-terminal end in the presence of pncRNA-D. A, an N- and C-terminal TLS-binding assay was performed under the following conditions. Lane 1, GST-tagged fragment 1 (N-terminal end) following PRMT1 and MNase treatment without adding fragment 4 (C-terminal end) was used as a negative control. Lane 2, fragment 4 was incubated with pncRNA-D and then precipitated with fragment 1. Lane 3, GST-tagged fragment 1 was precipitated with fragment 4. Lane 4, fragment 4 was treated with MNase and then incubated with fragment 1. Lane 5, MNase-treated fragment 4 was incubated with fragment 1 in the presence of pncRNA-D after the inhibition of the MNase activity by EGTA. Lane 6, MNase-treated, methylated fragment 4 was incubated with fragment 1. Lane 7, MNase-treated, methylated fragment 4 was incubated with fragment 1 in the presence of pncRNA-D after the inhibition of the MNase activity. Lane 8, 20% of the protein of fragment 4 was loaded as input. B, GST-TLS fragment 1 was precipitated with TLS fragment 4 (WT, R476A, and R481A) (lanes 2, 5, and 8), fragment 4 with MNase treatment (lanes 3, 6, and 9), and fragment 4 in the presence of pncRNA-D after inhibition of MNase activity (lanes 4, 7, and 10). Samples were stained with the SimplyBlueTM SafeStain for analysis.