Fig. 5: AZIN1 facilitates cell cycle progression in osteosarcoma cells in a polyamine-dependent manner.

A Representative gene set enrichment analysis (GSEA) results from RNA-seq data of 143B cells with or without AZIN1 knockdown, highlighting changes in cell cycle-related gene expression. B Gene expression heatmap displaying the transcriptional levels of cell cycle-regulating genes in 143B cells, comparing conditions with and without AZIN1 knockdown. C Representative flow cytometry plots of cell cycle analysis in 143B cells following AZIN1 knockdown and supplementation with 10 μM putrescine and 5 μM spermidine, assessed by BrdU pulse-labeling and PI staining. Quantification of cell cycle phase distribution is on the right. Data are mean ± SD (n = 3). Statistical significance was determined using one-way ANOVA. D Western blot analysis of cell cycle regulators in 143B cells with AZIN1 knockdown, supplemented with putrescine and spermidine. E In vitro growth curves of 143B cells with AZIN1 knockdown and subsequent supplementation with putrescine and spermidine. Data are mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA. F Normalized expression levels of MYC from RNA-seq data of 143B and U2-OS cells with and without AZIN1 knockdown. Data are mean ± SD (n = 3). Statistical significance was assessed using an unpaired t-test. G Representative IGV tracks showing MYC ChIP-seq binding on promoters of cell cycle-regulating genes in U2-OS cells, derived from the ChIP-Atlas database. Significance levels are indicated as follows: ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05, n.s. not significant.