Fig 2: Validation of key OSRDEGs expression at the mRNA levels.MAPK14, HIF1a, Myc, Hsp90ab1 and Hsp90aa1 mRNA level in the TAC and Sham groups in mice. Expression levels were standardized for GAPDH levels. A two-tailed unpaired Student’s t-test was used to compare two groups. Values are expressed as means ± SD (n = 4), *p < 0.05; **p < 0.01 and ***p < 0.001 vs Sham group.

Fig 3: Correlation analyzes between immune cell and hub genes.Association of MAPK14, HIF1a, Myc, Hsp90ab1 and Hsp90aa1 with 25 of types immune cells. Horizontal axis represents the immune cells; longitudinal coordinate represents the hub genes; The numbers in the red triangle represent correlation coefficient, the lager the number, the stronger the correlation. The numbers in the blue triangle represent p value.

Fig 4: Validation of key OSRDEGs expression at protein levels by western blotting.(A) Representative western blotting of MAPK14, HIF1a, Myc, Hsp90ab1 and Hsp90aa1 proteins of heart tissues in the TAC and Sham groups. (B) Quantitative analysis of MAPK14, HIF1a, Myc, Hsp90ab1 and Hsp90aa1 proteins of heart tissues in the TAC and sham groups. A two-tailed unpaired Student’s t-test was used to compare two groups. Values are expressed as means ± SD (n = 3). *p < 0.05 vs Sham group.

Fig 5: Hypoxia induces decreases in multimeric HSPs in human coronary arteries (HCAECs) and vein endothelial cells (HUVECs). (a) Whole-cell lysates from HCAECs exposed to hypoxia, heat-shock recovery, or basal conditions were subjected to native PAGE and sequentially stained for HSC70 (Invitrogen PA5-27337), HSP70 (Invitrogen MA3-009), HSP90 (CST 4877S), and HSP40 (CST 4868S), as shown in the top panels. Below, sodium dodecyl sulfate-PAGE results for each protein are displayed. Data are representative of n = 3 independent experiments using two different lots of cells (20TL365545 and 20TL064651, Lonza). Both native PAGE and Western blot experiments were performed using two different lots of cells (20TL365545 and 20TL064651). (b) Quantification of HSC70, HSP70, and HSP90 high-molecular-weight complexes in (a). Data are presented as the means ± SEMs, one-way ANOVA, with Tukey’s post hoc n = 3, *P < 0.05, **P < 0.01 versus basal conditions. Unpaired t test (#P < 0.05) was used to compare only the HSP90 hypoxic and basal groups. (c) Whole-cell lysates from HUVECs exposed to hypoxia or basal conditions were subjected to native PAGE and sequentially stained for HSC70 (Invitrogen PA5-27337) and HSP90 (CST 4877S), as shown in the top panels. Below, sodium dodecyl sulfate-PAGE for each protein is displayed. The data are representative of n = 2 independent experiments using cells from lot 19TL028324. (d) Whole-cell lysates of HUVECs exposed to hypoxia or basal conditions were subjected to Western blot analysis for HSP40 (CST 4868S), total HSP70 (Invitrogen MA3-006), HSC70 (Invitrogen PA5-27337), and HSP90 (CST 4877S). The data are representative of n = 3 independent experiments using cells from lots 19TL028324 and 23TL086130. (e) Data from (d) are presented as the mean ± SEM, unpaired Student’s t test, n = 3 with *P < 0.05 and ***P < 0.001. β-actin, loading control. (f) HUVECs subjected to basal or hypoxic 1% O2 (94% N2, 5% CO2) for 24 h were subjected to Western blot analysis for HIF1α (CST, #48085) with β-actin (Sigma, A5441) loading control. The assay was performed three times using two different lots of cells (21TL195720 and 23TL086130).