Fig 1: GRWD1 is important for maintaining the GRWD1-WDR5-MLL2 complex, but the three proteins also regulate distinct sets of genes. (A) GRWD1 knockdown reduced the efficiency of WDR5 immunoprecipitation of MLL2 but not histone H3. (B) GRWD1 overexpression increased the efficiency of WDR5 immunoprecipitation of MLL2 but not histone H3. (C and D) Heatmap of differential gene expression after GRWD1, WDR5, or MLL2 knockdown in MM (C) and KMM (D) cells. (E) Common and unique gene sets altered following GRWD1, WDR5, or MLL2 knockdown. (F) Shared and distinct common genes altered following GRWD1, WDR5, or MLL2 knockdown between MM and KMM cells. (G) The rank of the top enriched pathways of common genes altered following GRWD1, WDR5, or MLL2 knockdown in MM and KMM cells. NER, nucleotide excision repair. (H) RT-qPCR validation of CDK1 expression in MM and KMM cells after GRWD1, WDR5, or MLL2 knockdown. P values are from comparisons between each of the shRNA-treated groups (sh1, sh2, and sh3) and the scrambled control (Ctl).
Fig 2: GRWD1 directly interacts with WDR5. (A) Confocal images showing the colocalization of GRWD1 and WDR5 in MM and KMM cells. (B) FLAG-GRWD1 immunoprecipitated endogenous WDR5 in 293T cells. (C) FLAG-WDR5 immunoprecipitated endogenous GRWD1 in 293T cells. (D) Purified recombinant GST-GRWD1 but not GST physically pulled down purified WDR5 in vitro. (E) WDR5 knockdown reduced the levels of total H3K4me3 in MM and KMM cells.
Fig 3: WDR5 knockdown phenocopies GRWD1 knockdown. (A and B) Knockdown efficiencies of WDR5 shRNAs examined by RT-qPCR (A) and Western blotting (B). (C to E) The effects of WDR5 knockdown on cell proliferation (C), cell cycle progression (D), and apoptosis (E) in MM and KMM cells. (F) WDR5 knockdown reduced the efficiency of colony formation on soft agar of KMM cells. P values are from comparisons between each of the shRNA-treated groups (sh1, sh2, and sh3) and the scrambled control (Ctl).
Supplier Page from Abcam for Recombinant Human WDR5 protein (His tag N-Terminus)