Fig. 4: PLUM-EZH2 interaction is important for PRC2 complex formation. | Nature Communications

Fig. 4: PLUM-EZH2 interaction is important for PRC2 complex formation.

From: Multiple myeloma associated long non-coding RNA PLUM confers chemoresistance by enhancing PRC2 mediated UPR pathway activation

Fig. 4

a Levels of EED and SUZ12 protein co-immunoprecipitated by EZH2 antibody in scramble and sh2-PLUM KD KMS11 cells. (+MG132-20 µM). IgG: IP control; GAPDH: input protein loading control (N = 3 biological replicates). b Levels of EED and SUZ12 protein pulled down by FL-PLUM, ΔExon1 PLUM, ΔExon7 PLUM transcripts from nuclear lysate of scramble and sh-EZH2 KD KMS11 cells. IRE: negative control and GAPDH: input protein loading control (N = 1). c Levels of EZH2 and SUZ12 protein pulled down by FL-PLUM, ΔExon1 PLUM, ΔExon7 PLUM transcripts from nuclear lysate of scramble and sh-EED KD KMS11 cells. IRE: negative control and GAPDH: input protein loading control (N = 1). d Expression levels of EZH2 and EED protein pulled by FL-PLUM, ΔExon1 PLUM, ΔExon7 PLUM transcripts from nuclear lysate of scramble and sh-SUZ12 KD KMS11 cells. IRE: negative control and GAPDH: input protein loading control (N = 1). e Docked structure of exon7-PLUM and EZH2 protein. Interacting residues on EZH2 marked as Green: 488–498 aa; Red: 562–575 aa and Blue: 586–602 aa. f Graphical representation of the EZH2 protein with all the domains marked in different colors. (*) sign: reported residues having RNA-binding role. Mutants generated: HA-Mut1: K39A, HA-Mut2: F32AD36AK39A, HA-Mut3:F32AR34AD36AK39A and HA-Mut4: PRKKKR489-494NAAIRS. Tertiary structure of EZH2 protein with marked RNA-interacting residues in different color code. Green: F32-K39, Magenta: T345 (CDK1/2 phosphorylation site), Red: P489-R494 (disordered region). Created in BioRender. Deka, K. (2025) https://BioRender.com/f40k21z. g Levels of EZH2 protein pulled down with FL-PLUM in +/− CDK1 inhibitor (R03306) cells. DMSO: vehicle control and GAPDH: input protein loading control (N = 2 biological replicates). h Levels of wild type HA-EZH2 and HA-EZH2 mutants (HA-Mut1: K39A, HA-Mut2: F32AD36AK39A, HA-Mut3:F32AR34AD36AK39A and HA-Mut4: PRKKKR489-494NAAIRS) pulled down with FL-PLUM. IB: HA antibody and GAPDH: input protein loading control (N = 2 biological replicates). i Levels of EED, SUZ12 and EZH2 protein co-immunoprecipitated by HA antibody in wild type HA-EZH2 and HA-EZH2 mutants (HA-Mut1: K39A, HA-Mut2: F32AD36AK39A, HA-Mut3:F32AR34AD36AK39A and HA-Mut4: PRKKKR489-494NAAIRS) overexpressed KMS11 cells. IgG: IP control and GAPDH: input protein loading control (N = 2 biological replicates).

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