Fig 2: PKG regulates CHIP-chaperone interaction and post-translational stability.a Structural model of binding region between Hsc70 lid motif and the TPR region in CHIP. The location of S20 in reference to opposing lysine residues K597 and K601 in Hsc70 is shown. b Upper: example immunoprecipitation using Hsc70 as bait, and detecting Myc-tagged WT, SE, or SA CHIP in both control and SI conditions. There is minimal Hsc70-CHIP co-precipitation when CHIP-SA is expressed. Lower: summary data, n = 3 biological replicates/group, 1WANOVA for each condition, Holms-Sidaks mct: *p = 1.2 × 10−4, †0.003, ‡0.0005, §0.02. c Immunoprecipitation gels and summary data for Hsc70 and CHIP interaction in vivo with and without MI in mice with mPDE5oe and littermate controls (CON). Hsc70 is bait, and antibody detection using CHIP and Hsc70. N = 4 biological replicates/group, Kruskal-Wallis test, Dunns mct. P values in figure. Source data are provided as a Source Data file. Individual data and mean ± SEM shown in each summary panel.

Fig 3: Western blot showing APD-induced changes in the expression of the HIST1H4B, HSPA8, NCL, NPM1, and VIM proteins in C6 and B35 cells and the rat cortex. Nuclear protein extracts collected from APD-treated C6 cells, B35 cells and rat cortex were compared. a and (b), HIST1H4B; (c) and (d),HSPA8; (e) and (f), NCL; (g) and (h), NPM1; (i) and (j), VIM. Bar charts were constructed from triplicate western blot data obtained from three different batches of APD-treated cells or rat cortices using ANOVA followed by Dunnett’s test (*p-value < 0.05; **p-value < 0.01). Haloperidol (HAL), risperidone (RIS), clozapine (CLO), or PBS (control group, CTRL))

Fig 4: Verification of the expression of HSPA8 and DEK in HCC cells using qRT-PCR (A) and western blotting (B). The results are representative of three separate experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.

Fig 5: 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.