Figure 1.

Design of RNA/DNA Hybrid IP Method in HeLa Cells

(A) RNA/DNA hybrid IP workflow in HeLa cells.

(B) RNA/DNA hybrid slot blot with S9.6 antibody.

(C) Top: silver stain of RNA/DNA hybrid IP. No antibody and isotype-matched IgG2a antibody were used as controls. Bottom: western blot of RNA/DNA hybrid IP using indicated antibodies. Arrows indicate hypophosphorylated (IIa) and hyperphosphorylated (IIo) forms of Pol II. Triple amounts of input and IP samples were loaded for SETX.

(D) Top: silver stain of RNA/DNA hybrid IP following benzonase treatment. Bottom: western blot of RNA/DNA hybrid IP, probed with Top1 and H3 antibodies.

(E) Silver stain of RNA/DNA hybrid IP in the presence of RNA/DNA hybrid competitor.

(F) Western blot for Top1 of RNA/DNA hybrid IP with indicated synthetic competitors.

(C–E) ^∗, indicates the heavy chain of S9.6 and IgG2a antibodies. ^∗∗, indicates BSA, used to block protein A dynabeads.

See also Figure S1.