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. 2022 Jun 29;23(8):e54265. doi: 10.15252/embr.202154265

Figure 4. Vimentin protein level is upregulated by inhibition of PRMT5‐mediated sDMA.

Figure 4

  • A
    Immunofluorescence staining of V5‐vimentin wild‐type, 4RK and 4RF mutants expressed in CL1‐0 cells. F‐actin was stained with phalloidin, and nucleus was stained with DAPI. Scale bars, 10 μm. Data shown are representative of three independent experiments.
  • B
    Reconstitution of vimentin‐knockout cells with wild‐type, 4RK‐ and 4RF‐mutated V5‐vimentin. Both protein and mRNA levels of vimentin were determined by Western blots and RT‐qPCR. Data shown are representative of three independent experiments.
  • C
    Vimentin protein and mRNA levels in indicated paired cells were measured by Western blots and RT‐qPCR. Data shown are representative of three independent experiments.
  • D
    Effect of MTA on vimentin protein and mRNA levels determined by Western blots and RT‐qPCR. Data shown are representative of three independent experiments.
  • E
    Western blot analysis of vimentin protein levels in MTAP‐knockout and control cells transfected with PRMT5 siRNAs (left) or Flag‐PRMT5 (right). Arrows mark the site of Flag‐PRMT5 and endogenous PRMT5. Data shown are representative of three independent experiments.
  • F
    Effect of PRMT5 on the levels of ectopically expressed wild‐type, 4RK‐ and 4RF‐mutated V5‐vimentin in HEK293T cells assessed by Western blots. Data shown are representative of three independent experiments.
  • G
    Effects of wild‐type and catalytically dead R368A mutant PRMT5 on the level of V5‐vimentin in HEK293T cells detected by Western blot assays. Data shown are representative of three independent experiments.