Figure 6
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Figure 6
(A) The browser views showing the ATAC-seq peaks in keap1a promoter with in smarca5zko1049a and their siblings. Gray box indicates the change of ATAC-seq peaks after smarca5 deletion. The location of Gata1 motif at keap1a promoter is indicated by arrow. (B) qPCR analysis showing the expression of keap1a in RBCs from smarca5zko1049a and their siblings at 2 dpf. (C) qPCR analysis showing the expression of hmox1a, gclc, ggt1b, gsr, gstp1, gstk1, fbp1a, gsto2, prdx1, pgd and g6pd in RBCs from smarca5zko1049a and their siblings at 2 dpf. (D) The imaging of EGFP fluorescence in Tg (hsp70:keap1a-EGFP) embryos at 2 dpf. Heat shock was performed at 36 hpf. The bright-field of tail region in smarca5zko1049a and their siblings, with or without Smarca5 overexpression at 2 dpf. (E) The quantification of blood clots phenotype in (D). (F) The bright-field of tail region in smarca5zko1049a and their siblings, in control group and with hmox1a MO injection. The blood clots are indicated by arrow heads. (G) The quantification of blood clots phenotype in (F). (H) Schematic representation of Smarca5 in regulating erythrocyte aggregation via Keap1-Nrf2 signaling. In cytoplasm, Keap1 could anchor to Nrf2 to facilitate the Nrf2 degradation, while the release of Nrf2 to the nucleus could activate the expression of oxidation defense factors. In smarca5-deficient RBCs, the chromatin accessibility at keap1a promoters and the transcription of keap1a were decreased, which led to the excessive activation of hmox1a. Data are mean ± s.d. (B, C, E, G). Asterisk presents statistical significance (*p < 0.05, **p < 0.01, ***p < 0.001, n.s. not significant). p Values were calculated by two-tailed unpaired Student’s t-test.
Keap1-Nrf2 signaling pathway acts at downstream of Smarca5 in regulating RBC aggregation.