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A, Alignment of the SIN3A PRISMA binding profile to <t>ARID1A</t> tiled peptide array and the Sin3 SLiM ELM prediction on ARID1A . B, Bar graph representing levels of SIN3A on chromatin in ARID1A KO and parental RPE1 cells (n=3, mean ± SD, * p< 0.05, *** p< 0.00005). C, Co-immunoprecipitation of endogenous SIN3 complex (circled) by ARID1A C-terminal fragment GFP pulldown. D, Schematic of the in vitro binding experiment between <t>recombinant</t> <t>ARID1A-V5</t> and SIN3A fragment (449-1086) containing the PAH3 domain tagged with StrepII tag. E, Co-immunoprecipitation of recombinant ARID1A-V5 and SIN3A_PAH3-StrepII. ARID1A-V5 was immunoprecipitated from insect cell lysates. IgG was used as a negative control. Beads were then incubated with cell lysate from insect cells expressing SIN3A_PAH3-StrepII. Co-immunoprecipitation was assessed with anti-V5 and anti-StrepII antibodies to detect ARID1A and SIN3A_PAH3 respectively. KDM5A-StrepII was used as a positive control. Lanes labelled with X are irrelevant for this work.
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A, Alignment of the SIN3A PRISMA binding profile to <t>ARID1A</t> tiled peptide array and the Sin3 SLiM ELM prediction on ARID1A . B, Bar graph representing levels of SIN3A on chromatin in ARID1A KO and parental RPE1 cells (n=3, mean ± SD, * p< 0.05, *** p< 0.00005). C, Co-immunoprecipitation of endogenous SIN3 complex (circled) by ARID1A C-terminal fragment GFP pulldown. D, Schematic of the in vitro binding experiment between <t>recombinant</t> <t>ARID1A-V5</t> and SIN3A fragment (449-1086) containing the PAH3 domain tagged with StrepII tag. E, Co-immunoprecipitation of recombinant ARID1A-V5 and SIN3A_PAH3-StrepII. ARID1A-V5 was immunoprecipitated from insect cell lysates. IgG was used as a negative control. Beads were then incubated with cell lysate from insect cells expressing SIN3A_PAH3-StrepII. Co-immunoprecipitation was assessed with anti-V5 and anti-StrepII antibodies to detect ARID1A and SIN3A_PAH3 respectively. KDM5A-StrepII was used as a positive control. Lanes labelled with X are irrelevant for this work.
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A, Alignment of the SIN3A PRISMA binding profile to <t>ARID1A</t> tiled peptide array and the Sin3 SLiM ELM prediction on ARID1A . B, Bar graph representing levels of SIN3A on chromatin in ARID1A KO and parental RPE1 cells (n=3, mean ± SD, * p< 0.05, *** p< 0.00005). C, Co-immunoprecipitation of endogenous SIN3 complex (circled) by ARID1A C-terminal fragment GFP pulldown. D, Schematic of the in vitro binding experiment between <t>recombinant</t> <t>ARID1A-V5</t> and SIN3A fragment (449-1086) containing the PAH3 domain tagged with StrepII tag. E, Co-immunoprecipitation of recombinant ARID1A-V5 and SIN3A_PAH3-StrepII. ARID1A-V5 was immunoprecipitated from insect cell lysates. IgG was used as a negative control. Beads were then incubated with cell lysate from insect cells expressing SIN3A_PAH3-StrepII. Co-immunoprecipitation was assessed with anti-V5 and anti-StrepII antibodies to detect ARID1A and SIN3A_PAH3 respectively. KDM5A-StrepII was used as a positive control. Lanes labelled with X are irrelevant for this work.
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A, Alignment of the SIN3A PRISMA binding profile to <t>ARID1A</t> tiled peptide array and the Sin3 SLiM ELM prediction on ARID1A . B, Bar graph representing levels of SIN3A on chromatin in ARID1A KO and parental RPE1 cells (n=3, mean ± SD, * p< 0.05, *** p< 0.00005). C, Co-immunoprecipitation of endogenous SIN3 complex (circled) by ARID1A C-terminal fragment GFP pulldown. D, Schematic of the in vitro binding experiment between <t>recombinant</t> <t>ARID1A-V5</t> and SIN3A fragment (449-1086) containing the PAH3 domain tagged with StrepII tag. E, Co-immunoprecipitation of recombinant ARID1A-V5 and SIN3A_PAH3-StrepII. ARID1A-V5 was immunoprecipitated from insect cell lysates. IgG was used as a negative control. Beads were then incubated with cell lysate from insect cells expressing SIN3A_PAH3-StrepII. Co-immunoprecipitation was assessed with anti-V5 and anti-StrepII antibodies to detect ARID1A and SIN3A_PAH3 respectively. KDM5A-StrepII was used as a positive control. Lanes labelled with X are irrelevant for this work.
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A, Alignment of the SIN3A PRISMA binding profile to <t>ARID1A</t> tiled peptide array and the Sin3 SLiM ELM prediction on ARID1A . B, Bar graph representing levels of SIN3A on chromatin in ARID1A KO and parental RPE1 cells (n=3, mean ± SD, * p< 0.05, *** p< 0.00005). C, Co-immunoprecipitation of endogenous SIN3 complex (circled) by ARID1A C-terminal fragment GFP pulldown. D, Schematic of the in vitro binding experiment between <t>recombinant</t> <t>ARID1A-V5</t> and SIN3A fragment (449-1086) containing the PAH3 domain tagged with StrepII tag. E, Co-immunoprecipitation of recombinant ARID1A-V5 and SIN3A_PAH3-StrepII. ARID1A-V5 was immunoprecipitated from insect cell lysates. IgG was used as a negative control. Beads were then incubated with cell lysate from insect cells expressing SIN3A_PAH3-StrepII. Co-immunoprecipitation was assessed with anti-V5 and anti-StrepII antibodies to detect ARID1A and SIN3A_PAH3 respectively. KDM5A-StrepII was used as a positive control. Lanes labelled with X are irrelevant for this work.
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A, Alignment of the SIN3A PRISMA binding profile to ARID1A tiled peptide array and the Sin3 SLiM ELM prediction on ARID1A . B, Bar graph representing levels of SIN3A on chromatin in ARID1A KO and parental RPE1 cells (n=3, mean ± SD, * p< 0.05, *** p< 0.00005). C, Co-immunoprecipitation of endogenous SIN3 complex (circled) by ARID1A C-terminal fragment GFP pulldown. D, Schematic of the in vitro binding experiment between recombinant ARID1A-V5 and SIN3A fragment (449-1086) containing the PAH3 domain tagged with StrepII tag. E, Co-immunoprecipitation of recombinant ARID1A-V5 and SIN3A_PAH3-StrepII. ARID1A-V5 was immunoprecipitated from insect cell lysates. IgG was used as a negative control. Beads were then incubated with cell lysate from insect cells expressing SIN3A_PAH3-StrepII. Co-immunoprecipitation was assessed with anti-V5 and anti-StrepII antibodies to detect ARID1A and SIN3A_PAH3 respectively. KDM5A-StrepII was used as a positive control. Lanes labelled with X are irrelevant for this work.

Journal: bioRxiv

Article Title: High resolution interaction surface mapping by PRISMA reveals novel ARID1A interactions

doi: 10.64898/2026.03.15.711656

Figure Lengend Snippet: A, Alignment of the SIN3A PRISMA binding profile to ARID1A tiled peptide array and the Sin3 SLiM ELM prediction on ARID1A . B, Bar graph representing levels of SIN3A on chromatin in ARID1A KO and parental RPE1 cells (n=3, mean ± SD, * p< 0.05, *** p< 0.00005). C, Co-immunoprecipitation of endogenous SIN3 complex (circled) by ARID1A C-terminal fragment GFP pulldown. D, Schematic of the in vitro binding experiment between recombinant ARID1A-V5 and SIN3A fragment (449-1086) containing the PAH3 domain tagged with StrepII tag. E, Co-immunoprecipitation of recombinant ARID1A-V5 and SIN3A_PAH3-StrepII. ARID1A-V5 was immunoprecipitated from insect cell lysates. IgG was used as a negative control. Beads were then incubated with cell lysate from insect cells expressing SIN3A_PAH3-StrepII. Co-immunoprecipitation was assessed with anti-V5 and anti-StrepII antibodies to detect ARID1A and SIN3A_PAH3 respectively. KDM5A-StrepII was used as a positive control. Lanes labelled with X are irrelevant for this work.

Article Snippet: Codon-optimised human SIN3A (residues 449-1086) fused to StrepII tag and ARID1A-V5 (from Addgene plasmid #39311) cDNAs for insect cell expression were synthesised and subcloned into the pFastBac1 vector (GenScript).

Techniques: Binding Assay, Peptide Microarray, Immunoprecipitation, In Vitro, Recombinant, Negative Control, Incubation, Expressing, Positive Control