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. Author manuscript; available in PMC: 2022 Jun 22.
Published in final edited form as: Nat Immunol. 2021 Dec 22;23(1):99–108. doi: 10.1038/s41590-021-01087-w

Figure 2. TET deficiency is associated with increased levels of G-quadruplexes and R-loops.

Figure 2.

a) Diagrammatic representation of a G-quadruplex with an associated R-loop structure, illustrating the reagents used for detection of G-quadruplexes and R-loops. All experiments were performed in 8 to 10 week-old mice. b) Flow cytometric detection of G-quadruplexes by staining of permeabilized cells with BG4-Ig antibody or isotype IgG controls in B cells from CD19cre (YFP+), Dfl and CD19 DKO (YFP+) mice. c) Quantification of median fluorescence intensity (MFI) of BG4-Ig signal from CD19cre (YFP+), Dfl and CD19 DKO (YFP+) B cells from 4 independent experiments. d) Flow cytometric detection of G-quadruplexes after incubation of cells with NMM or DMSO vehicle controls (Veh) in B cells from CD19Cre (YFP+), Dfl and CD19 DKO (YFP+) mice. e) Quantification of median fluorescence intensity (MFI) of NMM signal from CD19Cre, Dfl and CD19 DKO B cells from 6 independent experiments. f) Flow cytometric detection of R-loops using V5-epitope-tagged recombinant RNASE H1 (rRNASE H1) in B cells from CD19Cre (YFP+), Dfl and CD19 DKO (YFP+) mice. Samples stained with anti-V5 and anti-rabbit secondary antibodies were used as controls (IgG). g) Quantification of median fluorescence intensity (MFI) of R-loops (rRNASE H1) signal from CD19cre (YFP+), Dfl and CD19 DKO (YFP+) B cells from 4 independent experiments. Statistical significance is calculated using one-way ANOVA in c), e) and g). Error bars represent mean +/− standard deviation, ** p value ≤0.01, *** p value≤0.002, **** p value ≤0.0001.