rabbit anti igf2bp3  (Santa Cruz Biotechnology)

Journal: Cell Discovery

Article Title: IGF2BP3-dependent glutamine/BCAA metabolic rewiring rejuvenates aged human adipose-derived stem cells for enhanced tissue regeneration

doi: 10.1038/s41421-025-00860-7

Figure Lengend Snippet: a Chord diagram showing that the highly expressed IGF2BP3 protein in I-hASCs is primarily involved in mRNA metabolism and stability pathways. b Differences in IGF2BP3 protein expression between I-hASCs and E-hASCs detected using iTRAQ and PRM. c Differential expression of IGF2BP3 mRNA between I-hASCs and E-hASCs using scRNA-seq. d Feature plots of IGF2BP3 mRNA expression in I-hASCs and E-hASCs based on the scRNA-seq data. e IGF2BP3 mRNA expression among the five hASC clusters, including I-ASCs and E-hASCs. f Deferential expression of IGF2BP3 mRNA between I-hASCs and E-hASCs in Cluster 1. g , h Pearson correlations of IGF2BP3 and BCAT1 mRNA expression ( g ) and of IGF2BP3 and GLS mRNA expression ( h ) in the total hASCs. i Western blotting assay was used to detect the protein expression of IGF2BP3, BCAT1, and GLS in I-hASCs after they were transfected with two distinct short hairpin RNA lentiviruses targeting IGF2BP3 (shIGF2BP3#1 and shIGF2BP3#2). j GSH, glutamate, and α-KG levels in I-hASCs with or without IGF2BP3 knockdown. k OCRs of I-hASCs with and without IGF2BP3 knockdown and quantification of the BR, AP, MR, and SC in each group. l , m Stability of BCAT1 ( l ) and GLS ( m ) mRNA in I-hASCs with or without IGF2BP3 knockdown. IGF2BP3 transcription was inhibited with actinomycin. n The m6A dot-blot assay revealed global RNA m6A levels after I-hASCs were transfected with two distinct short hairpin RNA lentiviruses targeting METTL3 (shMETTL3#1 and shMETTL3#2). Methylene blue staining served as a loading control. o m6A levels in I-hASCs with or without METTL3 knockdown. p Western blotting was used to detect METTL3, BCAT1, and GLS protein expression in I-hASCs after METTL3 knockdown. q qRT–PCR was used to measure BCAT1 (left panel) and GLS (right panel) mRNA expression levels in I-hASCs with or without METTL3 knockdown. r RIP assays in I-hASCs were used to assess the specific binding of IGF2BP3 to m6A-modified sites on BCAT1 mRNA (+1221 locus) and GLS mRNA (+4376 locus), and an anti-IGF2BP3-specific antibody was used for immunoprecipitation; an IgG antibody was used as a negative control. Analysis of MeRIP assays in I-hASCs to detect the recovery of BCAT1 mRNA +1221 locus ( s ) and GLS mRNA +4376 locus ( t ) with an anti-m6A antibody after METTL3 knockdown. u , v RIP assays were performed using an anti-IGF2BP3 antibody or IgG in I-hASCs after METTL3 knockdown. Luciferase activities of WT or Mut BCAT1 ( w ) and GLS ( x ) reporter vectors were quantified in I-hASCs under defined genetic perturbations: control, IGF2BP3 overexpression (IGF2BP3), or IGF2BP3 overexpression with METTL3 knockdown (IGF2BP3+shMETTL3). y IGF2BP3 was performed using streptavidin beads after S1m-WT/Mut-tagged BCAT1 and GLS mRNA were expressed. ** P < 0.01; *** P < 0.001.

Article Snippet: The primary antibodies used were anti-IGF2BP3 (#57145; 1:1000; Cell Signaling Technology), anti-BCAT1 (#88785; 1:1000; Cell Signaling Technology), and anti-GLS (#56750; 1:1000; Cell Signaling Technology) antibodies.

Techniques: Expressing, Multiplex sample analysis, Quantitative Proteomics, Western Blot, Transfection, shRNA, Knockdown, Dot Blot, Staining, Control, Quantitative RT-PCR, Binding Assay, Modification, Immunoprecipitation, Negative Control, Luciferase, Over Expression

Journal: Materials Today Bio

Article Title: 3D printing-biomimetic local stiff niche enhances glycolysis to boost PDAC cell stem-like phenotype via N6-methyladenosine-suppressed YAP1 mRNA decay

doi: 10.1016/j.mtbio.2025.102176

Figure Lengend Snippet: METTL14/IGF2BP3 respond to the high local stiffness suppressed the decay of YAP1 mRNA. (A – B) Predicted m6A motifs on YAP1 mRNA (SRAMP database, red: very high confidence, purple: high confidence). (C) Global m6A elevation in low, medium, and high local stiffness niches (Dot blot). (D) Overlap analysis of stiffness-responsive m6A WERs (writers, erasers, readers) targeting YAP1 by intersecting LOX and YAP1-correlated genes (R > 0.145, p < 0.05, TCGA-PAAD) and YAP1-predicted WERs (RM2Target). (E – I) The expression of METTL14, ZCCHC4, NOP58, IGF2BP2, and IGF2BP23 in PDAC tissues vs the normal (GEPIA2). (J) qRT-PCR analysis of METTL14, IGF2BP2, and IGF2BP3 in PDAC cells under different local stiffness niches (n = 3, p < 0.05). (K – N) Representative immunofluorescence staining (scale bar: 10 μm) and quantification showing increased METTL14 (red) and IGF2BP3 (violet) intensity of MIA-PACA2 cells in high local stiffness groups (n = 3, p < 0.05, vs low local stiffness group). (O – P) YAP1 mRNA stability assay of MIA-PACA2 cells in high local stiffness niches treated with Actinomycin D (10 μg/mL) after IGF2BP3 perturbation. (Q – R) Western blot results showing METTL14/IGF2BP3 knockdown or overexpression reciprocally regulate YAP1 protein levels of MIA-PACA2 cells in high local stiffness niches. (S) In MIA-PACA2 cells, the cooperative regulation of YAP1 by METTL14 and IGF2BP3 in high local stiff niches, β-actin served as control. Data: mean ± SD, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, and n.s.: denotes not significant. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Briefly, cells were lysed and incubated with magnetic beads conjugated with anti-rabbit METTL14 (5 μg, Proteintech, 26158-1-AP, China), anti-rabbit IGF2BP3 (5 μg, Proteintech, 14642-1-AP, China), or anti-IgG antibodies for more than 8 h at 4 °C.

Techniques: Dot Blot, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Stability Assay, Western Blot, Knockdown, Over Expression, Control

Journal: Materials Today Bio

Article Title: 3D printing-biomimetic local stiff niche enhances glycolysis to boost PDAC cell stem-like phenotype via N6-methyladenosine-suppressed YAP1 mRNA decay

doi: 10.1016/j.mtbio.2025.102176

Figure Lengend Snippet: Graphical illustration of the mechanism events involved in local high stiffness-regulated PDAC stemness. The PDAC TME is desmoplastic and mechanically heterogeneous, containing low and high local stiffness niches. Persistent stimuli in stiff local niches enhance the level of YAP1 via METTL14/IGF2BP3-suppressed YAP1 mRNA decay, leading to the augmentation of glycolysis and the acquisition of a stem-like phenotype for PDAC cells.

Article Snippet: Briefly, cells were lysed and incubated with magnetic beads conjugated with anti-rabbit METTL14 (5 μg, Proteintech, 26158-1-AP, China), anti-rabbit IGF2BP3 (5 μg, Proteintech, 14642-1-AP, China), or anti-IgG antibodies for more than 8 h at 4 °C.

Techniques: