prmt5 enzymatic activity  (MedChemExpress)

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: iScience

Article Title: m6A-dependent regulation of DGUOK-AS1 by RBM15 and HNRNPH1 in lung adenocarcinoma

doi: 10.1016/j.isci.2026.117078

Figure Lengend Snippet: DGUOK-AS1 201 and miR-2467-5p regulated PRMT5 expression (A) Predicted binding sites of miR-2467-5p on wild-type and mutant DGUOK-AS1 201 sequences. (B) Dual-luciferase reporter assay in A549 cells co-transfected with wild-type (WT) or mutant (MUT) DGUOK-AS1 201 and miR-2467-5p mimics ( n = 3, ∗∗ p < 0.01; ns, not significant). (C) RIP-qPCR analysis of DGUOK-AS1 enrichment on Ago2 relative to IgG in A549 cells ( n = 3, ∗∗∗ p < 0.001). (D) miR-2467-5p expression in BEAS-2B, A549, and H1975 cells was determined by RT-qPCR ( n = 3, ∗∗∗ p < 0.001). (E and F) miR-2467-5p levels in A549 and H1975 cells following DGUOK-AS1 knockdown or DGUOK-AS1 201 overexpression ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01). (G–J) Colony formation and Transwell assays assessing proliferation and migration of A549 cells co-transfected with DGUOK-AS1 201 overexpression vector and miR-2467-5p mimics (G and H: n = 3; I and J: scale bars, 100 μm; n = 5; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (K) Schematic of the predicted binding sites between miR-2467-5p and the wild-type or mutant 3′ UTR of PRMT5 mRNA. (L) Dual-luciferase reporter assay validating the interaction between miR-2467-5p and the PRMT5 3′ UTR in A549 cells ( n = 3, ∗∗∗ p < 0.001; ns, not significant). (M) RIP-qPCR analysis of PRMT5 mRNA enrichment using anti-Ago2 antibody or IgG control ( n = 3, ∗∗∗ p < 0.001). (N) PRMT5 protein expression in A549 and H1975 cells transfected with NC mimic or miR-2467-5p mimics. (O) PRMT5 protein levels in A549 and H1975 cells overexpressing DGUOK-AS1 201. (P and Q) CCK-8 and Transwell assays of proliferation and migration in A549 cells co-transfected with PRMT5 overexpression vector and DGUOK-AS1 siRNA (P: n = 3; Q: scale bars, 100 μm; n = 5; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). The data are shown as the mean ± SEM based on three independent trials. For (A–Q), n represents the number of independent biological replicates, unless otherwise specified. Statistical analysis was performed using a paired t test (C, E, F, and M), a one-way ANOVA (D, H, J, and Q), or two-way ANOVA (B and L), followed by Tukey’s multiple comparison test.

Article Snippet: Membranes were blocked using a solution of 5% nonfat dry milk and incubated with primary antibodies PRMT5 (1:800, A19533, ABclonal), RBM15 (1:1000, 60386, CST), HNRNPH1 (1:1000, 14774-1-AP, Proteintech), E-cadherin (1:1000, 20874-1-AP, Proteintech), Vimentin (1:1000, 10366-1-AP, Proteintech) and GAPDH (1:6000, AP0063, Bioworld), followed by reaction with HRP-conjugated secondary antibodies (1:4000, Proteintech).

Techniques: Expressing, Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Transfection, Quantitative RT-PCR, Knockdown, Over Expression, Migration, Plasmid Preparation, Control, CCK-8 Assay, Comparison

Journal: iScience

Article Title: m6A-dependent regulation of DGUOK-AS1 by RBM15 and HNRNPH1 in lung adenocarcinoma

doi: 10.1016/j.isci.2026.117078

Figure Lengend Snippet: RBM15 upregulates the expression of DGUOK-AS1 201 and promotes the proliferation and migration of LUAD cells (A–C) Tumor volume and weight in nude mice injected with A549 cells stably transfected with sh-RBM15 or sh-control ( n = 5 mice per group; ∗∗∗ p < 0.001). (D) Immunohistochemistry (IHC) staining of Ki67 in subcutaneous xenograft tumors (scale bars, 100 μm). (E and F) RT-qPCR analysis of DGUOK-AS1 expression in A549 and H1975 cells upon RBM15 knockdown or overexpression ( n = 3, ∗ p < 0.05, ∗∗∗ p < 0.001; ns, not significant). (G) Relative DGUOK-AS1 RNA levels measured by RT-qPCR following actinomycin D treatment ( n = 3, ∗ p < 0.05, ∗∗∗ p < 0.001). (H) Colony formation assay of proliferation in A549 cells co-transfected with RBM15 knockdown lentivirus and DGUOK-AS1 201 overexpression vector ( n = 3, ∗∗∗ p < 0.001). (I) Transwell migration assay in RBM15-silenced A549 cells overexpressing DGUOK-AS1 201 (scale bars, 100 μm; n = 5, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (J) RT-qPCR analysis of miR-2467-5p expression in A549 and H1975 cells upon RBM15 overexpression or knockdown ( n = 3, ∗ p < 0.05, ∗∗∗ p < 0.001). (K) Protein levels of RBM15 and PRMT5 in A549 cells with RBM15 overexpression or knockdown. Data are presented as mean ± SEM from three independent experiments. For (A–K), n represents the number of independent biological replicates, unless otherwise specified (mice: n = number of animals). Statistical analysis was performed using a paired t test (C and J), a one-way ANOVA (H and I), or a two-way ANOVA (E, F, and G) followed by Tukey’s multiple comparison test.

Article Snippet: Membranes were blocked using a solution of 5% nonfat dry milk and incubated with primary antibodies PRMT5 (1:800, A19533, ABclonal), RBM15 (1:1000, 60386, CST), HNRNPH1 (1:1000, 14774-1-AP, Proteintech), E-cadherin (1:1000, 20874-1-AP, Proteintech), Vimentin (1:1000, 10366-1-AP, Proteintech) and GAPDH (1:6000, AP0063, Bioworld), followed by reaction with HRP-conjugated secondary antibodies (1:4000, Proteintech).

Techniques: Expressing, Migration, Injection, Stable Transfection, Transfection, Control, Immunohistochemistry, Quantitative RT-PCR, Knockdown, Over Expression, Colony Assay, Plasmid Preparation, Transwell Migration Assay, Comparison

Journal: iScience

Article Title: m6A-dependent regulation of DGUOK-AS1 by RBM15 and HNRNPH1 in lung adenocarcinoma

doi: 10.1016/j.isci.2026.117078

Figure Lengend Snippet: HNRNPH1 regulates the expression of DGUOK-AS1 (A and B) RT-qPCR assay measuring the expression of DGUOK-AS1 following knockdown or overexpression of HNRNPH1 ( n = 3, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (C and D) The RNA stability of DGUOK-AS1 in A549 and H1975 cells was assessed after HNRNPH1 overexpression ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (E) The migration ability was measured using Transwell assay when DGUOK-AS1 201 was overexpressed in HNRNPH1-overexpression cells (scale bars, 100 μm; n = 4, ∗ p < 0.05). (F) CCK-8 assay was used to assess the proliferation of A549 cells co-transfected with HNRNPH1 and DGUOK-AS1 201 overexpression vectors ( n = 3, ∗∗∗ p < 0.001). (G) The expression of miR-2467-5p in A549 cells after overexpressing or knocking down HNRNPH1 by RT-qPCR analysis. (H) The protein levels of PRMT5 in A549 cells with overexpressed or knocked down HNRNPH1 ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01). (I) Molecular mechanism diagram of DGUOK-AS1 promoting cancer progression in LUAD. Each data point represents the mean ± SEM based on three independent trials. For (A–I), n represents the number of independent biological replicates, unless otherwise specified. Statistical analysis was performed using a paired t test (G), one-way ANOVA (E), or two-way ANOVA (A–D) and (F) followed by Tukey’s multiple comparison test.

Article Snippet: Membranes were blocked using a solution of 5% nonfat dry milk and incubated with primary antibodies PRMT5 (1:800, A19533, ABclonal), RBM15 (1:1000, 60386, CST), HNRNPH1 (1:1000, 14774-1-AP, Proteintech), E-cadherin (1:1000, 20874-1-AP, Proteintech), Vimentin (1:1000, 10366-1-AP, Proteintech) and GAPDH (1:6000, AP0063, Bioworld), followed by reaction with HRP-conjugated secondary antibodies (1:4000, Proteintech).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Over Expression, Migration, Transwell Assay, CCK-8 Assay, Transfection, Comparison

Journal: iScience

Article Title: m6A-dependent regulation of DGUOK-AS1 by RBM15 and HNRNPH1 in lung adenocarcinoma

doi: 10.1016/j.isci.2026.117078

Figure Lengend Snippet:

Article Snippet: Membranes were blocked using a solution of 5% nonfat dry milk and incubated with primary antibodies PRMT5 (1:800, A19533, ABclonal), RBM15 (1:1000, 60386, CST), HNRNPH1 (1:1000, 14774-1-AP, Proteintech), E-cadherin (1:1000, 20874-1-AP, Proteintech), Vimentin (1:1000, 10366-1-AP, Proteintech) and GAPDH (1:6000, AP0063, Bioworld), followed by reaction with HRP-conjugated secondary antibodies (1:4000, Proteintech).

Techniques: Virus, Recombinant, Transfection, RNA Extraction, Transwell Assay, Magnetic Beads, Lysis, Cell Counting, RNA Immunoprecipitation, Labeling, Silver Staining, Methylation, Immunoprecipitation, Immunohistochemistry, Luciferase, Reporter Assay, Mass Spectrometry, Software, RNA sequencing

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