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. 2024 Mar 21;44(4):469–490. doi: 10.1002/cac2.12534

FIGURE 4.

FIGURE 4

hnRNPA2B1 interacted with and stabilized NEAT1 in an m6A‐dependent manner. (A) Heatmap of differentially expressed lncRNAs (ǀlog2FCǀ > 1, P < 0.05, FPKM average > 0.5) in SGC7901ADR cells after hnRNPA2B1 knockdown. Red shades represent ADR_shNC, and blue shades represent ADR_sh. (B) Venn diagram of hnRNPA2B1‐binding lncRNAs. (C) hnRNPA2B1 knockdown or overexpression resulted in corresponding changes in NEAT1 levels. (D) qPCR analysis of NEAT1 expression in SGC7901 lv‐A2B1 and SGC7901VCR cells after METTL3 knockdown.(E) Analysis of MeRIP assays detecting NEAT1 retrieved by an m6A antibody in METTL3‐silenced SGC7901 lv‐A2B1 and SGC7901VCR cells. (F) Assessment of RIP assays detecting NEAT1 retrieved by an hnRNPA2B1 antibody or by IgG in SGC7901 lv‐A2B1 and SGC7901VCR cells. (G) Assessment of RIP assays detecting NEAT1 retrieved by an hnRNPA2B1 antibody or by IgG in METTL3‐silenced SGC7901 lv‐A2B1 and SGC7901VCR cells.(H) Assessment of the NEAT1 half‑life (t1/2) in hnRNPA2B1‑silenced SGC7901ADR cells. (I) Assessment of the NEAT1 half‑life (t1/2) in hnRNPA2B1‑silenced SGC7901VCR cells. (J‐K) IC50 values and growth curves of SGC7901 lv‐A2B1 cells transfected with ASO‐NEAT1 and treated with chemotherapy. The P value was calculated by one‐way ANOVA test (C left, D, H, and I), paired t test (C right and E, F, and G), one‐way ANOVA with Dunnett's multiple‐comparison test (H) and two‐way ANOVA with Dunnett's multiple‐comparison test (J and K). *P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant. Abbreviations: ADR_sh, hnRNPA2B1‐knockdown SGC7901ADR cell; ADR_shNC, negative control SGC7901ADR cell; ASO‐NC, negative control of antisense oligonucleotide; ASO‐NEAT1, antisense oligonucleotide of NEAT1; CRNDE, colorectal neoplasia differentially expressed; hnRNPA2B1, heterogeneous nuclear ribonucleoprotein A2B1; IgG, immunoglobulin G; IP, immunoprecipitation; lncRNA, long noncoding RNA; METTL3, methyltransferase 3; lv‐A2B1, overexpression of hnRNPA2B1; lv‐NC, empty overexpression control; MALAT1, metastasis associated lung adenocarcinoma transcript 1; NEAT1, nuclear enriched abundant transcript 1; shNC, negative control short hairpin RNA; shA2B1, short hairpin RNA of hnRNPA2B1.