Journal: Frontiers in Neurology

Article Title: PRMT5 inhibition suppresses the PI3K/AKT pathway to attenuate vascular smooth muscle cell pathological phenotype in intracranial aneurysm

doi: 10.3389/fneur.2026.1771196

Figure Lengend Snippet: PRMT5 level was increased in IA models. In vivo analysis of PRMT5 level between the sham and model groups (A) . In vitro analysis of VSMC viability (B) and PRMT5 level (C) among the untreated, Ang II (0.1 μM), Ang II (0.5 μM), and Ang II (1 μM) groups. *: p < 0.05; **: p < 0.01; ***: p < 0.001; ns: not significant.

Article Snippet: EPZ015666 (MCE, China) is an orally bioavailable small-molecule inhibitor of PRMT5 enzymatic activity.

Techniques: In Vivo, In Vitro

Journal: Frontiers in Neurology

Article Title: PRMT5 inhibition suppresses the PI3K/AKT pathway to attenuate vascular smooth muscle cell pathological phenotype in intracranial aneurysm

doi: 10.3389/fneur.2026.1771196

Figure Lengend Snippet: EPZ015666 inhibited PRMT5 enzymatic activity, facilitated transition from synthetic phenotype to contractile phenotype, and suppressed PI3K/AKT pathway in Ang II-treated VSMCs. Comparison of PRMT5, H4R3me2s, α -SMA, OPN, and MMP9 levels among the Model, EPZ (1 μM), EPZ (5 μM), and EPZ (10 μM) groups (A) . Comparison of p-PI3K and p-AKT among the Model, EPZ (1 μM), EPZ (5 μM), and EPZ (10 μM) groups (B) . *: p < 0.05; **: p < 0.01; ***: p < 0.001; ns: not significant.

Article Snippet: EPZ015666 (MCE, China) is an orally bioavailable small-molecule inhibitor of PRMT5 enzymatic activity.

Techniques: Activity Assay, Comparison

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: PRMT interacts with SHBs. (A) Huh7 cells were co–transfected with plasmids encoding SHBs–Strep–Flag (or Strep–Flag control) together with Flag–PRMT9. Strep pull–down was followed by WB with anti–Flag and anti–SHBs to assess co–precipitation. (B) Huh7 and HepG2 cells were co–transfected with plasmids encoding SHBs–Strep–Flag (or Strep–Flag control) together with Flag–PRMT5 and analyzed by Strep pull–down and WB as in (A). (C) Huh7 and HepG2 cells were co–transfected with plasmids encoding Strep–Flag–PRMT5 and SHBs–myc. Strep pull–down was performed and precipitates were immunoblotted for SHBs and Flag to validate the interaction. (D) Direct interaction between SHBs and PRMT5 was tested by GST pull–down. Purified GST or GST–PRMT5 (Coomassie–stained gel, left) was incubated with in vitro–translated SHBs–Flag, and bound SHBs was detected by WB using anti–Flag (right). (E) Confocal microscopy showing subcellular localization of SHBs (red) and PRMT5 (green) with nuclear DAPI staining (blue). Merged images and a representative line–scan fluorescence intensity profile (right) are shown.

Article Snippet: Cells were co–transfected with plasmids encoding Flag–PRMT5 together with SHBs–Strep–Flag or SHBs/R169K–Strep–Flag for 40 h. Cells were then treated with cycloheximide (CHX; 200 μg/mL; MedChemExpress, #HY–12320) to inhibit de novo protein synthesis.

Techniques: Transfection, Control, Purification, Staining, Incubation, In Vitro, Confocal Microscopy, Fluorescence

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: PRMT5 catalyzes symmetric dimethylation of SHBs at arginine 169. (A) Cells were transfected with plasmids encoding SHBs–Strep–Flag together with Flag–PRMT5 or control vector. Strep pull–down was followed by WB for SDMA and SHBs. Densitometric ratios (SDMA/IP–SHBs and SHBs/β–actin) are shown. (B) Cells expressing SHBs–Strep–Flag were transfected with control siRNA (NC) or two independent PRMT5 siRNAs. Strep pull–down/WB was used to assess SDMA on SHBs, and input blots confirmed PRMT5 knockdown and SHBs levels. (C) Cells expressing SHBs–Strep–Flag were treated with the PRMT5 inhibitor GSK591 (10 μM, 48 h) and analyzed by Strep pull–down/WB for SDMA and SHBs; densitometric ratios are shown. (D) Cells were transfected with plasmids encoding SHBs–Strep–Flag together with Flag–PRMT5 or Flag–PRMT5/E444Q. Strep pull–down was followed by WB for SDMA and SHBs. (E) Schematic of the in vitro methylation assay. Strep–Flag–PRMT5 and SHBs–Strep–Flag or SHBs/R169K–Strep–Flag were expressed in 293T cells, affinity–purified using streptavidin/Strep beads, eluted, and incubated ± SAM at 30 °C for 1.5 h prior to WB with SDMA antibody. (F) In vitro methylation reactions using the indicated purified proteins (WT SHBs or R169K mutant) incubated ± SAM. SDMA and total SHBs were detected by WB; SDMA/SHBs ratios are indicated above the blot.

Article Snippet: Cells were co–transfected with plasmids encoding Flag–PRMT5 together with SHBs–Strep–Flag or SHBs/R169K–Strep–Flag for 40 h. Cells were then treated with cycloheximide (CHX; 200 μg/mL; MedChemExpress, #HY–12320) to inhibit de novo protein synthesis.

Techniques: Transfection, Control, Plasmid Preparation, Expressing, Knockdown, In Vitro, Methylation, Affinity Purification, Incubation, Purification, Mutagenesis

Journal: Tumour Virus Research

Article Title: PRMT5–mediated symmetric dimethylation of SHBs at Arg169 stabilizes SHBs and promotes angiogenesis and tumor growth

doi: 10.1016/j.tvr.2026.200340

Figure Lengend Snippet: PRMT5 stabilizes SHBs protein expression in an Arg169–dependent manner. (A) Huh7 and HepG2 cells were co–transfected with plasmids encoding SHBs–Strep–Flag or SHBs/R169K–Strep–Flag together with increasing amounts of Flag–PRMT5 (0, 1, 3 μg). Whole–cell lysates were immunoblotted for SHBs, Flag, and β–actin; SHBs/β–actin ratios are shown above the blots. (B) Cells expressing SHBs–Strep–Flag or SHBs/R169K–Strep–Flag were transfected with NC or PRMT5 siRNAs (#1, #2). Lysates were immunoblotted for SHBs, PRMT5, and β–actin; SHBs/β–actin ratios are shown. (C–D) Cycloheximide (CHX) chase assays in (C) Huh7 and (D) HepG2 cells. Cells expressing SHBs or SHBs/R169K with vector or Flag–PRMT5 were treated with CHX for the indicated times (0–120 min), followed by WB for SHBs, Flag, and β–actin. Plots show relative SHBs levels normalized to time 0 with fitted linear regression (equations displayed). (E) HepG2 cells were co–transfected with plasmids encoding SHBs–Strep, HA–K48Ub, together with or without Flag–PRMT5, and treated with MG132 (20 μM) for 8 h, the ubiquitination levels of SHBs was evaluated via ubiquitination assay analysis. (F) HepG2 cells were co–transfected with plasmid encoding SHBs–Strep and TRIM21–myc (or control vector) and Flag–PRMT5 (or control vector), the cell lysates were subjected to immunoprecipitation using Strep–Tactin and analyzed by immunoblotting.

Article Snippet: Cells were co–transfected with plasmids encoding Flag–PRMT5 together with SHBs–Strep–Flag or SHBs/R169K–Strep–Flag for 40 h. Cells were then treated with cycloheximide (CHX; 200 μg/mL; MedChemExpress, #HY–12320) to inhibit de novo protein synthesis.

Techniques: Expressing, Transfection, Plasmid Preparation, Ubiquitin Proteomics, Control, Immunoprecipitation, Western Blot

Journal: bioRxiv

Article Title: PRMT5 as an Epigenetic Target for Group 3 (MYC-driven) Medulloblastoma

doi: 10.64898/2026.04.09.717536

Figure Lengend Snippet: (A) qPCR analysis for the expression of PRMT5/MYC mRNA in two MYC-amplified cell lines with transiently knocked-down of PRMT5 (using siRNAs) at 72 h. ****, p<0.0001 (student t test, SCR vs siPRMT5). (B) Effect of PRMT5 knockdown (siRNAs) on MYC-luciferase reporter gene (MYC-Luc) activity in HD-MB03 cells. *, p<0.05 (student t test) (C) ChIP analyses for the enrichment of PRMT5 and H4R3me2s on the proximal promoter region of the MYC gene in HD-MB03 cells. ****, p<0.0001 (student t test). ChIP analyses for the enrichment/binding of PRMT5 (D) and H4R3me2s (E) to the proximal promoter region of MYC gene, in PRMT5 knockdown (siRNAs) HD-MB03 cells. ( F ) Co-immunoprecipitation of BRD4 with PRMT5. ( G ) ChIP analyses for the enrichment/binding of PRMT5 and BRD4 to the proximal promoter region of MYC gene, in BRD4 knockdown (siRNAs) HD-MB03 cells. *, p<0.05; **, p<0.05 (student t test).

Article Snippet: Four PRMT5 inhibitors (EPZ015666, GSK332595, LLY-283, JNJ64619178) were purchased from MedChemExpress LLC or Selleckchem Company.

Techniques: Expressing, Amplification, Knockdown, Luciferase, Activity Assay, Binding Assay, Immunoprecipitation

Journal: bioRxiv

Article Title: PRMT5 as an Epigenetic Target for Group 3 (MYC-driven) Medulloblastoma

doi: 10.64898/2026.04.09.717536

Figure Lengend Snippet: RNA-sequencing was used to assess global gene expression changes in HD-MB03 cells 24 h after treatment with DMSO (vehicle) or JNJ64619178 (1 µM). (A) Volcano plot displaying genes significantly upregulated or downregulated in response to PRMT5 inhibition. (B) Gene Ontology Biological Process (GO-BP) functional analysis for top 10 pathways altered by JNJ. (C) Gene sets enrichment analysis (GSEA) (with p<0.01 and FDR<0.2) for top 10 pathways/gene sets (including MYC-associated target gene sets, RNA splicing, metabolism) altered by PRMT5 inhibition. (D) Number of the aberrant splicing events in protein coding genes disrupted by PRMT5 inhibition. (E) GO-BP functional analysis for the aberrant splicing events altered by PRMT5 inhibition. (F) Volcano plot represents all the significant (p<0.05) splicing events. Selected genes from the indicated GO-BP functional classification are highlighted. Metabolism associated genes GGT6, GPT2 and C1QBP1 are identified.

Article Snippet: Four PRMT5 inhibitors (EPZ015666, GSK332595, LLY-283, JNJ64619178) were purchased from MedChemExpress LLC or Selleckchem Company.

Techniques: RNA Sequencing, Gene Expression, Inhibition, Functional Assay

Journal: bioRxiv

Article Title: PRMT5 as an Epigenetic Target for Group 3 (MYC-driven) Medulloblastoma

doi: 10.64898/2026.04.09.717536

Figure Lengend Snippet: (A) MTT assay showing the effects of PRMT5 inhibitors (1-50 µM) on cell growth of MYC-amplified MB (HD-MB03, D341), non-MYC MB (ONS-76) and normal human astrocyte (NHA) cell lines. (B) IC 50 values of PRMT5 inhibitors in the indicated cell lines. (C) Annexin-V assay showing effects of PRMT5 inhibitors (JNJ, EPZ) on apoptosis in HD-MB03 cells. **, p<0.01, ***, p<0.001 (relative to ‘0’ (vehicle control)). (D) Cell cycle profile in HD-MB03 cells treated with PRMT5 inhibitors (EPZ, JNJ). (E) Western blot analysis for the expression of the indicated key proteins in JNJ-treated HD-MB03 cells. ( F) Quantification of spheres following treatment of JNJ and EPZ in two PDX-derived MB cell lines (MED-411FH, MED-114FH) at 72 h. Values, mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.005; *** p < 0.001 (Student- t -test). (G) Representative sphere (MED-411FH) images showing disruption of spheres in each treatment. (H) Western blot results showing the expression of indicated proteins in MED-411 spheres treated with JNJ and EPZ.

Article Snippet: Four PRMT5 inhibitors (EPZ015666, GSK332595, LLY-283, JNJ64619178) were purchased from MedChemExpress LLC or Selleckchem Company.

Techniques: MTT Assay, Amplification, Annexin V Assay, Control, Western Blot, Expressing, Derivative Assay, Disruption

Journal: bioRxiv

Article Title: PRMT5 as an Epigenetic Target for Group 3 (MYC-driven) Medulloblastoma

doi: 10.64898/2026.04.09.717536

Figure Lengend Snippet: (A ) JNJ brain concentration in BALB/c mice (N=3) following oral administration of 10 mg/kg JNJ at different timepoints. **p<0.01 (Student’s t-test). (B) NSG mice (N=5) with subcutaneously xenografted HD-MB03 cells were treated orally with vehicle or JNJ (10 mg/kg) five times a week for three weeks. Tumor volume measurement of xenografted mice following treatments. The differences noted between treatment groups show comparison by Student ‘s t-test of the tumor volumes on 21 days post treatment (p<0.005). (C) Representative IHC images (40 × magnification with 60 µm scale bar) of PRMT5, MYC, Ki-67, and CC3 expression in xenografts 21 days post-treatment, as indicated. Bar graphs below show the percentages of PRMT5, MYC, Ki-67 and CC3 positive cells derived from immunohistology scores, which were semi-quantitated in the tumors of three xenografted mice. **p<0.01; ***p<0.005 (Student’s t-test). (D) NSG mice (N=6) with orthotopically xenografted HD-MB03 cells were treated daily with vehicle or JNJ (10 mg/kg) or JNJ (F) (10 mg/kg) for two weeks. Survival analysis of xenografted mice using Kaplan-Meier (long-rank test). *p<0.05, **p<0.01, ***p<0.001. (E) Representative IHC images (4x magnification with 600 µm scale bar) and respective quantification showing MYC-positive tumors in the mouse cerebellum. The percentage of MYC, derived from immunohistology scores, was semi-quantitated in the tumors of three xenografted mice 21 days post-treatment. *p< **p<0.01 (Student’s t-test).

Article Snippet: Four PRMT5 inhibitors (EPZ015666, GSK332595, LLY-283, JNJ64619178) were purchased from MedChemExpress LLC or Selleckchem Company.

Techniques: Concentration Assay, Comparison, Expressing, Derivative Assay