Fig. 2. Significant up-regulation of β/γ-crystallins was found in human with high myopia.

a Heat map from gene expression microarray analysis of up-regulated and down-regulated genes in lens epithelium of emmetropic control and highly myopic eyes showing 117 differentially expressed genes (n = 3 vs. 3, fold change ≥ 2, all p < 0.05, CRYBB1 p = 0.009, CRYGD p = 0.032, CRYBA2 p = 0.024, CRYBA4 p = 0.022, CRYBA1 p = 0.032). Right, the top 15 up-regulated genes. b qPCR analyses of mRNA expression of the five β/γ-crystallin genes (n = 3, CRYBB1 p = 2.11e−5, CRYGD p = 0.0005, CRYBA2 p = 0.0003, CRYBA4 p = 0.0002, CRYBA1 p = 0.0001). c Examination of protein expression by Western blotting of the five β/γ-crystallin genes (n = 4, CRYBB1 p = 0.002, CRYGD p = 0.007, CRYBA2 p = 0.007, CRYBA4 p = 0.021, CRYBA1 p = 0.019). Right, the band density was normalized to loading control as a ratio for statistical analysis. d Immunofluorescence images of CRYBB1 and CRYBA2 staining in the human lens epithelium (n = 7). Scale bar: 50 μm. e Correlation between the increased mRNA and protein levels of the five target β/γ-crystallins in lens epithelium and the lens equatorial diameter in highly myopic eyes (examined by qPCR and PRM). n=biological replicates. In (a) and (d), one specimen from an individual was used as one sample; in (b, c, and e), pooled samples were used, of which in (e), lens epithelial samples were pooled in order of lens equatorial diameter (three pieces of lens epithelial samples pooled as one and totally 48 pieces used here) and the average lens equatorial diameter of each three pooled samples were used in the scatterplot. Results are presented as mean ± SD. Level of significance was detected using two-sided Student’s t test (a–c). Associations between variables were evaluated by Pearson correlation analysis (e), two-sided test was used). ****p < 0.0001, ***p < 0.001, **p < 0.01, and *p < 0.05. Source data are provided as a Source Data file.