Figure S3. BAZ1B domain targeting PCH.... | Rockefeller University Press

Figure S3.

$BAZ1B domain targeting PCH. (A) Schematic drawing of various truncated BAZ1B and their localizations. Images of BAZ1B-FL and BrD+AT1-3 are reproductions of those in Fig. 6 A. Contrast was adjusted individually to compare the localization patterns. (B) Expression and purification of EGFP-tagged BAZ1B BrD+AT1-3 and BrD+AT1-3mut were evaluated by SDS-polyacrylamide gel electrophoresis and Coomassie Brilliant Blue staining. HEK293T cells transfected with the expression vectors were lysed and the supernatant (input) was incubated with GFP Trap anti-GFP nanobody magnetic beads at 4°C overnight. After removing the unbound fraction (Flowthrough), beads were washed, and aliquots were mixed with SDS-gel loading buffer (GFP-IP). Serial dilution series of BSA were loaded to estimate the amounts of recovered protein bound to beads. (C–E) Nucleotide sequences of DNA used for binding assay. (C) Major satellite repeats. (D) EF1α promoter. (E) 601 DNA. Sequences with more than three consecutive AT-bases are highlighted in red. Scale bar, 10 μm. Source data are available for this figure: SourceData FS3.$

BAZ1B domain targeting PCH. (A) Schematic drawing of various truncated BAZ1B and their localizations. Images of BAZ1B-FL and BrD+AT1-3 are reproductions of those in Fig. 6 A. Contrast was adjusted individually to compare the localization patterns. (B) Expression and purification of EGFP-tagged BAZ1B BrD+AT1-3 and BrD+AT1-3mut were evaluated by SDS-polyacrylamide gel electrophoresis and Coomassie Brilliant Blue staining. HEK293T cells transfected with the expression vectors were lysed and the supernatant (input) was incubated with GFP Trap anti-GFP nanobody magnetic beads at 4°C overnight. After removing the unbound fraction (Flowthrough), beads were washed, and aliquots were mixed with SDS-gel loading buffer (GFP-IP). Serial dilution series of BSA were loaded to estimate the amounts of recovered protein bound to beads. (C–E) Nucleotide sequences of DNA used for binding assay. (C) Major satellite repeats. (D) EF1α promoter. (E) 601 DNA. Sequences with more than three consecutive AT-bases are highlighted in red. Scale bar, 10 μm. Source data are available for this figure: SourceData FS3.

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