Journal: iScience

Article Title: circIFNGR2 regulating ankylosing spondylitis-associated inflammation through macrophage polarization

doi: 10.1016/j.isci.2023.107325

Figure Lengend Snippet: eIF4A3 regulated the expression of circIFNGR2 (A) The binding sites of eIF4A3 in the flanking sequences of the IFNGR2 pre-mRNA transcript were predicted by circInteractome. (B) RIP assays for detection of the binding relationships among eIF4A3 and IFNGR2 pre-mRNA transcripts. (C) Western Blot analysis for identification of the binding relationships among eIF4A3 and the flanking sequences of IFNGR2 pre-mRNA transcripts. (D) qRT-PCR analysis of the expression level of eIF4A3 mRNA in THP-1 cells, M0 macrophages, M1 macrophages, and M2 macrophages. (E) qRT-PCR analysis of the expression level of eIF4A3 mRNA, circIFNGR2, and linear IFNGR2 with or without overexpression and knockdown of eIF4A3. (F) qRT-PCR analysis of the expression of circIFNGR2 in PBMCs from 24 healthy controls and 33 patients with AS. (G) Correlation analysis of the expression levels of circIFNGR2 and eIF4A3 mRNA in PBMCs from patients with AS. Data in B, D, and E are presented as mean ± SD, data in F are presented as median and quartiles. ∗∗: p < 0.01; ∗∗∗: p ＜0.001; ns: p ≥ 0.05.

Article Snippet: Rabbit polyclonal anti-eIF4A3 , Bioss , Cat# bs-14548R.

Techniques: Expressing, Binding Assay, Western Blot, Quantitative RT-PCR, Over Expression, Knockdown

Journal: iScience

Article Title: circIFNGR2 regulating ankylosing spondylitis-associated inflammation through macrophage polarization

doi: 10.1016/j.isci.2023.107325

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-eIF4A3 , Bioss , Cat# bs-14548R.

Techniques: Virus, Recombinant, Adjuvant, In Vivo, SYBR Green Assay, Transfection, Lysis, Blocking Assay, Western Blot, DNA Extraction, Protein Extraction, Enzyme-linked Immunosorbent Assay, Purification, RNA Immunoprecipitation, Luciferase, Software

Journal: RNA

Article Title: The RNA-binding profile of Acinus, a peripheral component of the exon junction complex, reveals its role in splicing regulation

doi: 10.1261/rna.057158.116

Figure Lengend Snippet: Proportion of Acinus peaks associated with eIF4A3 peaks and enrichment at “canonical” EJC occupancy site (24-nt upstream of an exon junction). ( A ) Proportion of Acinus peaks found in close proximity of eIF4A3 peaks (eIF4A3+) (maximum distance of 10 nt between both peak summits). The analysis was done on “genomic” peaks (reads mapped to the genome) depending on their location: all peaks ( n = 36,271), exonic peaks ( n = 25,347), 5′UTR peaks ( n = 1347), 3′UTR peaks ( n = 2379), and intronic peaks ( n = 6944), but also on “transcriptomic” peaks (reads mapped to a representative transcriptome data set) ( n = 13,231). eIF4A3 CLIP-seq data described in were downloaded from GEO: GSM1001331. ( B ) Distribution of the peak frequency according to the distance of the peak summit from the exon 5′ end or the exon 3′ end. This analysis was done using Acinus’ “common transcriptomic” peak data for all peaks ( n = 10,735), peaks associated with eIF4A3 ( n = 2000), or peaks not associated with eIF4A3 ( n = 8735) (from left to right ). Only internal exons were used. The iCLIP data are represented by a red line. The gray line corresponds to randomized peak summit positions.

Article Snippet: The following commercial antibodies were used: anti-Acinus antibody (Calbiochem, ab-2 PC552), anti-T7 mouse monoclonal (Novagen, 69522), anti-eIF4A3 antibody (Proteintech, 17504-1-AP), and anti-β-tubulin (Sigma, T4026).

Techniques:

Journal: Biology of Reproduction

Article Title: Nuclear localization of EIF4G3 suggests a role for the XY body in translational regulation during spermatogenesis in mice †

doi: 10.1093/biolre/iox150

Figure Lengend Snippet: Primary antibodies used in this study.

Article Snippet: 1:100 n.a Thermo EIF4E Mo Thermo MA1–089 AB_2536738 1:100 1:2000 EIF4E2 Mo Santa Cruz sc-100731 AB_1122491 1:100 n/a EIF4E3 Rab ProteinTech 17282–1-AP AB_2262162 1:100 n/a EIF4A1 Mo Thermo MA5–14901 AB_11006110 1:100 n/a EIF4A2 Rab ProteinTech 11280–1-AP AB_2097378 1:100 1:2000 EIF4A3 Rab Abcam Ab32485 AB_732124 1:50 n/a SYCP3 Rat Handel Lab. Eaker et al.,2001 n.a.

Techniques:

Journal: Molecular Cancer

Article Title: Long noncoding RNA ZFAS1 promoting small nucleolar RNA-mediated 2′-O-methylation via NOP58 recruitment in colorectal cancer

doi: 10.1186/s12943-020-01201-w

Figure Lengend Snippet: SNORD12C/78-mediated 2′-O-Me modification regulates target genes expression in a ZFAS1-dependent manner. a and b , EIF4A3 and LAMC2 mRNA expression was determined after knocking down SNORD12C or SNORD78 by qRT-PCR assays( a ), and Western blot method( b ). c , Reducing EIF4A3 and LAMC2 half-life by silencing SNORD12C treated by the ActD in SW620 cells. d , Reducing the translation activity of EIF4A3 and LAMC2 by silencing SNORD12C and SNORD78 treated by the CHX in SW620 cells. e , The 28S rRNA G3878 and G4593 2′-O-Me activities were declined after silencing SNORD12C or SNORD78 expression in SW620 and HCT116 cells detected by RTL-P assays. f , Overexpressing ZFAS1 rescued the 2′-O-Me activities under the lower dNTPs conditions after silencing SNORD12C and SNORD78. g, The statistical plot of overexpressing ZFAS1 rescued the 2′-O-Me activities after silencing SNORD12C and SNORD78 *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

Article Snippet: Membranes were immunoblotted with anti-rabbit NOP58 (1:1000), EIF4A3 (1:1000) (Proteintech, Chicago, USA; Abcam, UK) and anti-mouse LAMC2 (1:500), GAPDH (1:2000) (Abcam, UK; Zsbio, Beijing, China), and then were incubated with hybrid secondary antibody, and the data was collected by FluorChem V2.0 (Alpha Innotech Corp, USA).

Techniques: Modification, Expressing, Quantitative RT-PCR, Western Blot, Activity Assay

Journal: Molecular Cancer

Article Title: Long noncoding RNA ZFAS1 promoting small nucleolar RNA-mediated 2′-O-methylation via NOP58 recruitment in colorectal cancer

doi: 10.1186/s12943-020-01201-w

Figure Lengend Snippet: ZFAS1 inhibited proliferation by targeting NOP58 protein in vivo. a , Schematic diagram of xenografts in BALB/c nude mice by inoculating HCT116 cells that were stably co-transfected with ZFAS1, ZFAS1-NOP58-Wild, and ZFAS1-NOP58-Mut, as well as the control with empty vector at their right armpits. Then half of the xenografts were sacrificed at the 35th day after injection and the other half were tracked until death. b , Mean tumor weight of each group xenografts in nude mice. Data are shown as mean ± s.d., n = 6 for each group. c , Mean tumor volumes on different days for each group xenografts in nude mice. Data are showed as mean ± s.d., n = 6 for each group. d , Representative tumors size excised on day 35 are shown. e , Kaplan-Meier graph showing overall survival of each group, n = 6. f , qPCR assays were performed to determine the (m) RNA expression of ZFAS1, NOP58, LAMC2, EIF4A3, SNORD12C, SNORD78 in above each group. g, h and i , IHC assay and western blot to detect the protein expression of NOP58 in xenografts tumor tissues of each group. The groups were as follows: NC (empty vector); ZFAS1 (pcDH-ZFAS1); ZFAS1 + NOP58-Wild (co-transfected with pcDH-ZFAS1 and pcDH-NOP58-Wild); ZFAS1 + NOP58-Mut (co-transfected with pcDH-ZFAS1 and pcDH-NOP58-Mut).*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

Article Snippet: Membranes were immunoblotted with anti-rabbit NOP58 (1:1000), EIF4A3 (1:1000) (Proteintech, Chicago, USA; Abcam, UK) and anti-mouse LAMC2 (1:500), GAPDH (1:2000) (Abcam, UK; Zsbio, Beijing, China), and then were incubated with hybrid secondary antibody, and the data was collected by FluorChem V2.0 (Alpha Innotech Corp, USA).

Techniques: In Vivo, Stable Transfection, Transfection, Control, Plasmid Preparation, Injection, RNA Expression, Western Blot, Expressing

Journal: Molecular Cell

Article Title: RNA Polymerase II Phosphorylated on CTD Serine 5 Interacts with the Spliceosome during Co-transcriptional Splicing

doi: 10.1016/j.molcel.2018.09.004

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-eIF4A3 , Bethyl , Cat# 302-981A-T; RRID: AB_10748369.

Techniques: Sequencing, Mass Spectrometry, Software