Journal: Cellular and molecular life sciences : CMLS

Article Title: IGF2BP1-HAX-1 positive feedback loop-mediated HAX-1 overexpression blocks autophagic flux and promotes chemoresistance in nasopharyngeal carcinoma.

doi: 10.1007/s00018-025-05604-0

Figure Lengend Snippet: Figure 8 Schematic overview of the role of HAX-1 in DDP chemoresistance by effectively blocking autophagic flux in NPC. IGF2BP1 enhanced HAX-1 m6A methylation, thereby enhancing its stability. Furthermore, HAX-1 binds IGF2BP1 mRNA, thereby contributing to its stability and translation. Moreover, HAX-1 recruits NEDD4 to promote Rab7a degradation and inhibits binding of Rab7a with SNAREs by competitively binding to it, thereby blocking the fusion of autophagosomes with lys- osomes

Article Snippet: Co-IP co-immunoprecipitation, WB western blot ◂ Reagent or Resource Source Identifier IGF2BP1 proteintech 22803-1-AP NEDD4 proteintech 21698-1-AP HA-Tag Cell Signaling Tech- nology #3724 Myc-Tag Cell Signaling Technology #2276 YTHDF1 proteintech 17479-1-AP YTHDF2 proteintech 24744-1-AP WTAP proteintech 60188-1-Ig GAPDH proteintech 10494-1-AP Bacterial and Virus Strains lentiviral HAX1- shRNA Shanghai Genechem Target seq: GAG TGA TGC AAG AAG TGA A lentiviral IGF2BP1shRNA Shanghai Genechem Target seq: ACA GTA GAG AAC TGT GAG CAA lentiviral Rab7AshRNA Shanghai Genechem Target seq: gaAAC AAG ATT GAC CTC GAA A RFP-GFP-tagged LC3 Shanghai Genechem tfLC3 Biological Samples Mouse tissues This paper N/A Human NPC speci- mens Affiliated Hospital of Nantong University N/A Chemicals, peptides, and recombinant proteins DMSO Fisher Scientific Cat#BP231-100 Cisplatin Jiangsu HANSOH PHARMA H20040813 Hochest Thermo Fisher Scientific 62249 Propidium Iodide Becton Dickinson and Company 556463 Fetal Bovine Serum Biological Industries 04-001-1ACS Reagent or Resource Source Identifier RPMI 1640 Biological Industries 01–100-1ACS TRIzol reagent BBI B511311 Uranyl acetate Polysciences, Hirschberg an der Bergstrasse, Germany 6159-44-0 Basic lead citrate Sigma-Aldrich, Taufkirchen, Germany 15326 Actinomycin D MCE, Sollentuna, Sweden HY-17559 Cycloheximide MCE, Sollentuna, Sweden HY-12320 MG-132 MEC, Sollentuna, Sweden HY-13259 AR6 buffer AKOYA, Marlborough, Massachusetts, USA AR600 CQ, chloroquine MCE HY-17589A Critical commercial assays EdU cell prolifera- tion assay Shanghai Beyotime C0078S CCK8 kit BBI E606335 LIVE/DEAD viabil- ity kit Shanghai Bestbio BB-4126 BCA protein assay kit Thermo Fisher Scientific 23227 Annexin V-FITC Apoptosis Detection Kit Beyotime C1063 Dual-Luciferase Reporter Assay Kit Beyotime RG088M Oligonucleotides Primer1 for HAX1, Forward CAG GAG GAG GGA TAC GTT TCC This paper N/A Primer1 for HAX1, Reverse CCC ATA TCG CTG AAG ATG CTATT This paper N/A Primer1 for preHAX1, Forward ATT ACA GTC GCC TGC CAA AC This paper N/A Primer1 for preHAX1, Reverse AGG TCA GGA GCT CAA AAG CA This paper N/A Primer1 for IGF2BP1, Forward GCG GCC AGT TCT TGG TCA A This paper N/A Primer1 for IGF2BP1, Reverse TTG GGC ACC GAA TGT TCA ATC This paper N/A Figure 5 Regulation of IGF2BP1 mRNA stability and translation by IGF2BP1.

Techniques: Blocking Assay, Methylation, Binding Assay