Figure 2

Figure 2 Validation of lncRNA expression changes in CMS and non-CMS cells under hypoxia and normoxia.

(A) qRT-PCR validation of all 5 lncRNAs that were differentially altered (up- and downregulated) in the CMS cell group (Supplemental Table 1). iPSC-derived CD34⁺ cells after exposure to hypoxia and normoxia (for 3 days) were used for this assay. Expression levels were tested and validated in both CMS (n = 3) and non-CMS (n = 3) cells under hypoxia and normoxia. *P < 0.05, t test. HIKER/LINC02228 was tremendously upregulated in the CMS cells under hypoxia. (B) qRT-PCR validation of top 10 upregulated lncRNAs in the non-CMS cell group (Supplemental Table 1). iPSC-derived CD34⁺ cells after exposure to hypoxia and normoxia (for 3 days) were used for this assay. Expression levels were tested and validated in both CMS (n = 3) and non-CMS (n = 3) cells under hypoxia and normoxia. *P < 0.05, t test. (C) qRT-PCR validation of top 10 downregulated lncRNAs in the non-CMS cell group (Supplemental Table 1). iPSC-derived CD34⁺ cells after exposure to hypoxia and normoxia (for 3 days) were used for this assay. Expression levels were tested and validated in both CMS (n = 3) and non-CMS (n = 3) cells under hypoxia and normoxia. *P < 0.05, t test. (D) Nuclear and cytoplasmic localization of lncRNAs. qRT-PCR results of confirmation for the expression changes for HIKER/LINC02228, LINC00431 (nuclear) and LINC01133, and APOBEC3B-AS1 and UBE2Q1-AS1 (cytoplasmic) for CMS, non-CMS and sea-level erythroid cells under hypoxia and normoxia. iPSC-derived CD34⁺ cells after exposure to hypoxia and normoxia (for 3 days) were used for this assay. *P < 0.05, t test. n = 3 subjects for each group.