Fig 2: Screening of immune-related histone chaperones and clustering of 369 TCGA-HCC samples based on HSPA8 and DEK. (A) Correlation analysis between 36 known histone chaperones and the four ESTIMATE indices, including stromal score, immune score, ESTIMATE score and tumor purity. (B,C) Comparison of the mRNA expression of HSPA8 or DEK between tumor and normal tissues from the TCGA database. (D,E) Comparison of the protein level of HSPA8 or DEK between tumor and normal tissues from the HPA database. Scale bar, 100 µm. (F,G) Correlation analysis between HSPA8 or DEK and the abundance of tumor-infiltrating lymphocytes by TISIDB. (H) TCGA-HCC samples are divided into two clusters according to HSPA8 and DEK via consensus clustering. (I) Spearman rank correlation analysis between the expression of HSPA8 and DEK. *** p < 0.001.

Fig 3: DEK regulated DRP1 via ATAD3A. ChIP-Seq was performed on the control (Input) and DEK overexpressing BEAS-2B cells (HA). (A) Peak density histograms following ChIP-Seq. (B) Heat map of peak density. (C) Statistics of the distribution of the peak region, such as 3′ UTR, 5′ UTR, Distal Intergenic, Downstream, Exon, Intron, Promoter, etc. (D) Gene Ontology enrichment analysis after screening using the keyword of mitochondria. (E) Peak density of ATAD3A. (F) ChIP-quantitative real-time PCR analysis of ATAD3A mRNA levels. (G, H) Western blot analysis and quantification of ATAD3A in lung tissue of DEK wild type (WT) and DEK knockout (KO) mice pretreated with HDM. (I) The expression of ATAD3A mRNA level and (J) DRP1 mRNA level in lung tissue. Western blot analysis of ATAD3A, DRP1. and DEK after co-immunoprecipitation in BEAS-2B cells (K–N). Western blot analysis of ATAD3A and DRP1 after co-immunoprecipitation in BEAS-2B cells (O, P) and lung tissues (Q, R). Data were presented as mean ± SEM. n=3. *p<0.05, **p<0.01, ***p<0.001.

Fig 4: Activation effects of DEK on mitophagy, NLRP3 inflammasome, and apoptosis in BEAS-2B cells. BEAS-2B cells were pretreated with 3-MA (5 mM) for 4 h and then incubated with rmDEK (1 μg/ml) for 24 h. (A) Mitochondrial morphology was observed by transmission electron microscopy. Scale bar = 2 μm. (B–E) Western blot analysis and quantification of COX IV, SQSTM1, LC3B I/II, PINK1, Parkin, and MnSOD. (F, G) Representative images and quantification of TUNEL staining in BEAS-2B cells. Scale bar = 1000 μm. (H–K) Western blot analysis and quantification of caspase-1, NLRP3, IL-1β, Cleaved caspase-3, Bax, and Bcl-2. Data were presented as mean ± SEM. n=3. *p<0.05, **p<0.01, ***p<0.001.

Fig 5: miR-5100 inhibits autophagy and promotes apoptosis in GC cells. (A) miR-5100 mimics/mimics-NC were transfected into HGC27 or SGC7901 cell lines stably expressing mCherry-EGFP-LC3B. Cells were treated with 5 nM rapamycin (Rap) 24 h after transfection, and autophagic flux was observed 8 h later (scar bar = 40 μm). (B) Autophagosome (red dots) and Autolysosome (yellow dots) were counted and analyzed, **: p < 0.01. (C) miR-5100 mimics/mimics-NCs were transfected into HGC27 or SGC7901 cell lines. Cells were treated with 5 nM rapamycin (Rap) 24 h after transfection, and the level of autophagy was observed 8 h later. (D,E) miR-5100 mimics/mimics-NCs were transfected into HGC27 or SGC7901 cell lines, and cells were harvested 36 h later. Western Blot detection of DEK, p62, LC3B proteins. The ratio of LC3BII/I was analyzed by normalization with Actin (n = 3, **: p < 0.01). (F,G) miR-5100 mimics/mimics-NC were transfected into HGC27 or SGC7901 cell lines, and the apoptosis rate was analyzed by flow cytometry after 36 h, and statistical analysis was performed (n = 3, **: p < 0.01, ***: p <0.001). (H,I) miR-5100 mimics/mimics-NC was transfected into HGC27 or SGC7901 cell lines, and 36 h later, the expression of Cleaved-caspase-3 was detected by Western Blot, and quantitative analysis was performed (n = 3, **: p < 0.01, ***: p <0.001).