prmt5 recombinant protein  (Sino Biological)

Journal: Scientific reports

Article Title: Discovery of an ApoE4-targeted small-molecule SirT1 enhancer for the treatment of Alzheimer's disease.

doi: 10.1038/s41598-025-96131-2

Figure Lengend Snippet: Fig. 1. DDL-214 racemate and enantiomer effects on target mRNA and proteins. (A) The structure and molecular weight of DDL-218. Levels of SirT1 (B) protein (n = 6) and (C) mRNA (n = 5 for DDL-214 and 218; n = 6 for DDL-219) in murine N2a-E4 cells after treatment with DDL-214 (racemate), or DDL-218(-) enantiomer or DDL-219(+) enantiomer at 5 µM for 24 h. (D) Levels of NFYb, PRMT5, SirT1, and PPP2r5e mRNA in human Kelly neuroblastoma cells treated with DDL-214, DDL-218, or DDL-219 (n = 8). Data normalized to DMSO control. Data graphed as the mean and SEM. Statistical analysis performed for (B) and (D) using one-way ANOVA and Tukey’s post-hoc comparisons, where*p ≤ .05 and ****p < .000.1 An unpaired student’s t-test was performed for the comparison in (C) (one-way ANOVA was not significant). mRNA data are normalized to GAPDH mRNA in the same samples and to levels in DMSO (veh)-treated cells.

Article Snippet: Cells were then Scientific Reports | (2025) 15:14028 2| https://doi.org/10.1038/s41598-025-96131-2 transfected with 0.75 μg of pCMV6-Entry vector (Origene # PS100001) or the human ORF clone for PRMT1 (Origene # RC224239), PRMT4 (Origene # RC217483), PRMT8 (Origene # RC205188), PRMT5 (Origene # RC203458), or PRMT7 (Origene # RC201672) using TurboFectin transfection reagent (Origene# TF81001) following manufacturer’s protocol.

Techniques: Molecular Weight, Control, Comparison

Journal: Scientific reports

Article Title: Discovery of an ApoE4-targeted small-molecule SirT1 enhancer for the treatment of Alzheimer's disease.

doi: 10.1038/s41598-025-96131-2

Figure Lengend Snippet: Fig. 2. PRMT5 overexpression increases, and knockdown decreases, SirT1 expression. (A) SirT1 mRNA levels after transfection of N2a-E4 cells with either empty pCMV vector or a PRMT5-expressing vector (over- expression) (n = 16). (B) Fold change in SirT1 protein level after PRMT5 vs. empty vector transfection. (C) The levels of SirT1 and PRMT5 mRNA normalized to GAPDH, and GAPDH mRNA levels (not normalized) after transfection of N2a-E4 with vehicle (untreated), scrambled siRNA or PRMT5 siRNA (n = 9). Statistical analysis performed using an unpaired, two-tailed student’s t-test where **p < .01 and ***p < .001. Comparison in (B) is between scrambled siRNA and PRMT5 siRNA.

Article Snippet: Cells were then Scientific Reports | (2025) 15:14028 2| https://doi.org/10.1038/s41598-025-96131-2 transfected with 0.75 μg of pCMV6-Entry vector (Origene # PS100001) or the human ORF clone for PRMT1 (Origene # RC224239), PRMT4 (Origene # RC217483), PRMT8 (Origene # RC205188), PRMT5 (Origene # RC203458), or PRMT7 (Origene # RC201672) using TurboFectin transfection reagent (Origene# TF81001) following manufacturer’s protocol.

Techniques: Over Expression, Knockdown, Expressing, Transfection, Plasmid Preparation, Two Tailed Test, Comparison

Journal: Scientific reports

Article Title: Discovery of an ApoE4-targeted small-molecule SirT1 enhancer for the treatment of Alzheimer's disease.

doi: 10.1038/s41598-025-96131-2

Figure Lengend Snippet: Fig. 3. Barnes maze latency, gene mRNA levels, and SirT1 promoter occupancy. (A) Latency in seconds (sec) for mice to reach escape hole in the 48-hour probe in the Barnes maze. Mice who were immobile are not represented. (B) Gene mRNA levels for NFYb, PRMT5, and SirT1 in hippocampus of study mice, the color code in (A) also applies to (B). (C) Occupancy of the SirT1 promoter by RNAP or ApoE4 in brain tissue of ApoE4-TR:5xFAD mice treated with either vehicle-only (Veh) or DDL-218. Data graphed as mean and SEM. Statistical analysis performed using one-way ANOVA with Tukey’s post-hoc comparison of groups in (A) and by unpaired two-tailed student’s t-test for (B) and (C), where *p ≤ .05, for comparison of ApoE4-5XFAD Veh vs. DDL-218 mice in (A) and between RNAP promoter binding in Veh vs. DDL-218 in (C).

Article Snippet: Cells were then Scientific Reports | (2025) 15:14028 2| https://doi.org/10.1038/s41598-025-96131-2 transfected with 0.75 μg of pCMV6-Entry vector (Origene # PS100001) or the human ORF clone for PRMT1 (Origene # RC224239), PRMT4 (Origene # RC217483), PRMT8 (Origene # RC205188), PRMT5 (Origene # RC203458), or PRMT7 (Origene # RC201672) using TurboFectin transfection reagent (Origene# TF81001) following manufacturer’s protocol.

Techniques: Comparison, Two Tailed Test, Binding Assay

Journal: Scientific reports

Article Title: Discovery of an ApoE4-targeted small-molecule SirT1 enhancer for the treatment of Alzheimer's disease.

doi: 10.1038/s41598-025-96131-2

Figure Lengend Snippet: Fig. 4. Proposed mechanism of action for SirT1 enhancement by DDL-218. (1) ApoE4 binds the SIRT1 promoter CLEAR sequence, acting as a transcriptional brake (repressor) and reducing SirT1 expression. (2) Small molecule DDL-218 increases expression of transcription factor NFYb, which then leads to (3) increased PRMT5 expression. (4) PRMT5 then binds ApoE4, decreasing ApoE4 occupancy of the SirT1 promoter, results in de-repression of SirT1 promoter or ‘release of the brake’. (5) This allows for increased RNA polymerase binding to the SIRT1 promoter, and increased SirT1 expression. The 3D protein images of ApoE4, NFYb, PRMT5 and RNA Pol were generated using Cresset software from PDB: 2L7B (ApoE4), 8QU3 (NFYb), 4GQB (PRMT5), 4A45 (RNA Pol).

Article Snippet: Cells were then Scientific Reports | (2025) 15:14028 2| https://doi.org/10.1038/s41598-025-96131-2 transfected with 0.75 μg of pCMV6-Entry vector (Origene # PS100001) or the human ORF clone for PRMT1 (Origene # RC224239), PRMT4 (Origene # RC217483), PRMT8 (Origene # RC205188), PRMT5 (Origene # RC203458), or PRMT7 (Origene # RC201672) using TurboFectin transfection reagent (Origene# TF81001) following manufacturer’s protocol.

Techniques: Sequencing, Expressing, Binding Assay, Generated, Software