Journal: Cell reports

Article Title: SART3 reads methylarginine-marked glycine- and arginine-rich motifs

doi: 10.1016/j.celrep.2024.114459

Figure Lengend Snippet:

Article Snippet: SART3 , Bethyl , Cat# A301–521A; RRID: AB_999668.

Techniques: Virus, Recombinant, Protease Inhibitor, Membrane, Mutagenesis, SYBR Green Assay, cDNA Synthesis, Knock-Out, Plasmid Preparation, Software

Journal: Cell reports

Article Title: SART3 reads methylarginine-marked glycine- and arginine-rich motifs.

doi: 10.1016/j.celrep.2024.114459

Figure Lengend Snippet: Figure 1. The HAT repeats of SART3 read SDMA-marked GAR motifs (A) Schematic representation of the GFP-Trap-based IP-mass spectrometry (MS) approach. (B) List of GFP-RMET interacting proteins identified by IP-MS. HEK293T cells were transfected with GFP-RMET, and half the cells were treated with AdOx for 3 days and subjected to IP-MS analysis. The number of peptides identified for each protein is listed.

Article Snippet: SART3 interacts with a methylated arginine motif A GFP-Trap approach (developed by ChromoTek), using nanobody-based reagents, is an efficient way to identify protein-protein interactions in cells.30 Here, we used a GFP-Trap to identify additional effectors of methylarginine marks.

Techniques: Mass Spectrometry, Protein-Protein interactions, Transfection

Journal: Cell reports

Article Title: SART3 reads methylarginine-marked glycine- and arginine-rich motifs.

doi: 10.1016/j.celrep.2024.114459

Figure Lengend Snippet: Figure 1. The HAT repeats of SART3 read SDMA-marked GAR motifs (A) Schematic representation of the GFP-Trap-based IP-mass spectrometry (MS) approach. (B) List of GFP-RMET interacting proteins identified by IP-MS. HEK293T cells were transfected with GFP-RMET, and half the cells were treated with AdOx for 3 days and subjected to IP-MS analysis. The number of peptides identified for each protein is listed.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies SART3 Bethyl Cat# A301-521A; RRID: AB_999668 GFP (WB) Santa Cruz Biotechnology Cat# sc-9996; RRID: AB_627695 GFP (IP) Invitrogen Cat# A6455; RRID: AB_2314549 GST This paper N/A Fibrillarin Proteintech Cat# 16021-1-AP; RRID: AB_2105788 Coilin Cell Signaling Technology Cat# 14168; RRID: AB_2798410 Actin Sigma-Aldrich Cat# A1978; RRID: AB_476692 PABP(ADMA) Cell Signaling Technology Cat# 3505; RRID: AB_2298971 FAM(ADMA) In house N/A ADMA In house N/A SDMA In house N/A MMA In house N/A PRMT1 Bethyl Cat# A300-722A; RRID: AB_533421 PRMT5 Active Motif Cat# 61001 SND1 Bethyl Cat# A302-883A; RRID: AB_10631268 SMN Bethyl Cat# A301-862A; RRID: AB_1309784 EFTUD2 Proteintech Cat# 10208-1-AP; RRID: AB_2095834 SNRNP200 Proteintech Cat# 23875-1-AP; RRID: AB_2879346 SART1 Proteintech Cat# 22675-1-AP; RRID: AB_2879148 PRPF6 Proteintech Cat# 23929-1-AP; RRID: AB_2879365 AGO1 Cell Signaling Technology Cat# 5053; RRID: AB_2616013 SNRPB Santa Cruz Biotechnology Cat# sc-271094; RRID: AB_10611786 La Bethyl Cat# A303-902A; RRID: AB_2620252 USP4 Bethyl Cat# A300-829A; RRID: AB_597888 USP15 Bethyl Cat# A300-923A; RRID: AB_2214726 Peroxidase AffiniPure Donkey Anti-Mouse IgG Jackson ImmunoResearch Cat# 715-035-151; RRID: AB_2340771 Peroxidase AffiniPure Donkey Anti-Rabbit IgG Jackson ImmunoResearch Cat# 711-035-152; RRID: AB_10015282 Bacterial and virus strains E. coli Top10 Thermo Fisher Scientific Cat# C404010 E. coli BL21 Thermo Fisher Scientific Cat# C600003 Chemicals, peptides, and recombinant proteins DMEM Medium Corning Cat# MT10017CV Penicillin-Streptomycin Corning Cat# MT30002CI Fetal bovine serum Thermo Fisher Scientific Cat# 10500056 PBS Corning Cat# MT21031CV Trypsin-EDTA 0.25% Corning Cat# MT25053CI Blasticidin S Thermo Fisher Scientific Cat# A1113903 Puromycin Thermo Fisher Scientific Cat# A1113802 Complete Protease Inhibitor Cocktail Sigma-Aldrich Cat# 4693159001 Phosphatase Inhibitor Cocktail Thermo Fisher Scientific Cat# 78420 Phenylmethylsulfonyl fluoride (PMSF) Sigma-Aldrich Cat# P7626 GFP-Trap kit Chromotek Cat# gtak-20 NP-40 Sigma-Aldrich Cat# I8896 DMSO Sigma-Aldrich Cat# D5879 (Continued on next page) Cell Reports 43, 114459, July 23, 2024 11

Techniques: Mass Spectrometry, Protein-Protein interactions, Transfection

Journal: Aging

Article Title: SART3, regulated by p53, is a biomarker for diagnosis, prognosis and immune infiltration in hepatocellular carcinoma.

doi: 10.18632/aging.204978

Figure Lengend Snippet: Figure 2. High expression of SART3 correlates with poor prognosis and advanced staging of HCC. Kaplan-Meier curves show SART3high and SART3low groups for (A) OS, (B) DSS and (C) PFS in TCGA dataset, and for (D) OS and (E) RFS in GSE14520 dataset. Violin plot shows SART3 expression at different AFP levels (F), T-stage (G), histological grade (H) and pathological stage (I) expression levels, *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: In this study, a polyclonal antibody targeting SART3 (Proteintech, Wuhan, China) was used at a concentration of 1:400.

Techniques: Expressing

Journal: Aging

Article Title: SART3, regulated by p53, is a biomarker for diagnosis, prognosis and immune infiltration in hepatocellular carcinoma.

doi: 10.18632/aging.204978

Figure Lengend Snippet: Figure 3. Prognostic value of SART3 in HCC. (A) Univariate Cox regression model and (B) multivariate Cox regression model in TCGA. (C) Univariate Cox regression model and (D) Multivariate Cox regression model in GSE14520. Kaplan-Meier survival curves for subgroups of SART3high and SART3low patients in (E) female, (F) male, (G) T1 and T2, (H) T3 and T4, (I) AFP ≤400, (J) AFP >400, (K) stage I and II and (L) stage III and IV.

Article Snippet: In this study, a polyclonal antibody targeting SART3 (Proteintech, Wuhan, China) was used at a concentration of 1:400.

Techniques:

Journal: Aging

Article Title: SART3, regulated by p53, is a biomarker for diagnosis, prognosis and immune infiltration in hepatocellular carcinoma.

doi: 10.18632/aging.204978

Figure Lengend Snippet: Figure 6. High expression of SART3 upregulates the expression level of immune checkpoint genes. (A) Heat map showing co- expression of SART3 and immune checkpoint genes. (B) Correlation matrix showing SART3-immune checkpoint association. (C) Expression levels of immune checkpoint genes between SART3 high and SART3 low groups. (D) Correlation of SART3 with immune checkpoint genes was verified in TIMER database.

Article Snippet: In this study, a polyclonal antibody targeting SART3 (Proteintech, Wuhan, China) was used at a concentration of 1:400.

Techniques: Expressing

Journal: Aging

Article Title: SART3, regulated by p53, is a biomarker for diagnosis, prognosis and immune infiltration in hepatocellular carcinoma.

doi: 10.18632/aging.204978

Figure Lengend Snippet: Figure 7. Identification and functional annotation of DEGs and GSEA analysis. (A) Volcano plot showing DEGs between SART3 high and SART3 low groups. (B) Pie chart showing the percentage of up-regulated DEGs and down-regulated DEGs in transcriptome genes. (C) Heat map showing the top 25 up- and down-regulated genes. (D) Bubble plots of DEGs significantly enriched in GO terms and KEGG pathways. Enrichment plots showing (E) four significantly positively correlated KEGG pathways and (F) four significantly positively correlated GO annotations.

Article Snippet: In this study, a polyclonal antibody targeting SART3 (Proteintech, Wuhan, China) was used at a concentration of 1:400.

Techniques: Functional Assay

Journal: Aging

Article Title: SART3, regulated by p53, is a biomarker for diagnosis, prognosis and immune infiltration in hepatocellular carcinoma.

doi: 10.18632/aging.204978

Figure Lengend Snippet: Figure 9. SART3 and its related genes are closely associated with RNA transcription in HCC cells. (A) PPI network of SART3- associated genes. (B) Annotations and correlation coefficients of nine SART3-related genes. (C, D) Enriched GO and KEGG pathways of SART3 and its related genes.

Article Snippet: In this study, a polyclonal antibody targeting SART3 (Proteintech, Wuhan, China) was used at a concentration of 1:400.

Techniques:

Journal: Aging

Article Title: SART3, regulated by p53, is a biomarker for diagnosis, prognosis and immune infiltration in hepatocellular carcinoma.

doi: 10.18632/aging.204978

Figure Lengend Snippet: Figure 10. Representative images and corresponding histograms of the wound healing assay of Huh-7. (A) and SUN-449 (B) cells in si-SART3 group and control group. Representative images (C) and corresponding histograms (D, E) of the Trans-well assay of Huh-7 and SUN-449 cells in si-SART3 group and control group. Growth curves of Huh-7 (F) and SUN-449 (G) cells in si-SART3 group and control group by CCK-8 assay.

Article Snippet: In this study, a polyclonal antibody targeting SART3 (Proteintech, Wuhan, China) was used at a concentration of 1:400.

Techniques: Wound Healing Assay, Control, CCK-8 Assay