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. 2020 Nov 6;2(4):zcaa032. doi: 10.1093/narcan/zcaa032

Figure 2.

Figure 2.

CX-5461 inhibits initiation but not recruitment of RPI–Rrn3. (A) Graphical representation of qPCR amplicons within the mouse rDNA repeat unit. (B, C) ChIP-qPCR analysis, respectively, of RPI and Rrn3 interactions across the rDNA unit of NIH3T3 cells after 15 min exposure to either 2 or 10 μM CX-5461 or mock treated by the addition of vehicle (NaH2PO4). Standard errors were <0.1 and <0.03 for RPI and Rrn3, respectively, and so have been omitted. (D) DChIP-seq profiles of RPI and Rrn3 interactions across the NIH3T3 rDNA unit after treatment with 10 μM CX-5461, 40 nM ActD or mock treatment for 15 min. The lower panel shows an enlargement of the region boxed in the upper panel. The key elements of the rDNA repeat are indicated graphically below both panels. The scale of ChIP enrichment in (D) is indicated at the upper right of each lane. In (A) to (D), SpPr refers to the Spacer promoter [yellow in (D)]; Tsp, Spacer promoter adjacent terminator [red in (D)]; T0, 47S promoter proximal terminator; 47SPr, 47S pre-rRNA promoter; 47S 5′, amplicon covering 5′ base pairs 159–320 of the 47S pre-rRNA coding region; ETS, external transcribed sequence; 18S, 5.8S and 28S indicate structural rRNA coding regions; T1–10, 47S termination elements; and T1–3, the qPCR amplicon for this region.