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To assess the role of SETX on R-loop formation within the IgH locus, we isolated R-loops from whole <t>DNA</t> and probed the IgH locus for their prevalence. (A) A not-to-scale representative image of the mouse IgM and IgA regions within the IgH locus, approximate probe locations are marked. DNA was isolated from <t>whole</t> <t>splenocytes</t> and R-loops were immunoprecipitated following 24 hours of (B) no treatment, (C) polyclonal stimulation, and (D) IgA specific stimulation. (E) RNA was isolated from splenocytes treated with IgA stimulation for 72 hours and the expression levels of AID, IgM-GLT, and IgA-GLT were assessed via q-PCR (n = 3). Statistics were determined via two-way ANOVA with Bonferroni post-hoc test, p < 0.001, ***.
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To assess the role of SETX on R-loop formation within the IgH locus, we isolated R-loops from whole DNA and probed the IgH locus for their prevalence. (A) A not-to-scale representative image of the mouse IgM and IgA regions within the IgH locus, approximate probe locations are marked. DNA was isolated from whole splenocytes and R-loops were immunoprecipitated following 24 hours of (B) no treatment, (C) polyclonal stimulation, and (D) IgA specific stimulation. (E) RNA was isolated from splenocytes treated with IgA stimulation for 72 hours and the expression levels of AID, IgM-GLT, and IgA-GLT were assessed via q-PCR (n = 3). Statistics were determined via two-way ANOVA with Bonferroni post-hoc test, p < 0.001, ***.

Journal: bioRxiv

Article Title: Senataxin is required for optimal class switch recombination to the immunoglobulin A isotype

doi: 10.1101/2025.09.23.678005

Figure Lengend Snippet: To assess the role of SETX on R-loop formation within the IgH locus, we isolated R-loops from whole DNA and probed the IgH locus for their prevalence. (A) A not-to-scale representative image of the mouse IgM and IgA regions within the IgH locus, approximate probe locations are marked. DNA was isolated from whole splenocytes and R-loops were immunoprecipitated following 24 hours of (B) no treatment, (C) polyclonal stimulation, and (D) IgA specific stimulation. (E) RNA was isolated from splenocytes treated with IgA stimulation for 72 hours and the expression levels of AID, IgM-GLT, and IgA-GLT were assessed via q-PCR (n = 3). Statistics were determined via two-way ANOVA with Bonferroni post-hoc test, p < 0.001, ***.

Article Snippet: RNA was isolated from stimulated mouse splenocytes using the RNeasy isolation kit (QIAgen) and 1 μg of complimentary DNA was synthesized using the Maxima First Strand cDNA Synthesis kit (ThermoFisher Scientific) according to the manufacturer’s instructions.

Techniques: Isolation, Immunoprecipitation, Expressing