Figure 6.
Figure 6. Altered expression of select erythroid differentiation and proliferation genes upon reduction of TR4 activity in human and murine erythroid cells. (A,B) RNA-seq analysis from CD34+ cells infected with a sh-TR4 lentivirus, as previously reported,36 and RNA-seq results from control jumbled shRNA (shaded bars) or the shRNA-TR4 (white bars) values are represented as fragments per kilobase per million reads (FKPM).36 (C) qRT-PCR quantification of multiple genes expressed in erythroid cells of stage III (CD71+TER119+) cells sorted from both WT and Tr4+/− mouse total bone marrow. The fold changes were calculated using ∆∆Ct with WT set to 1 (indicated by the dashed line). (D) Schematic summarizing the known functions of TR4 in murine erythroid cells, including these data. Dashed lines indicate correlative data indicating Cdkn1c, Gata1, Alas2, Alad, and Gata2 are TR4 candidate target genes regulating erythroid proliferation and maturation. Solid arrows from TR4 to εy-globin and βh1-globin indicate direct transcriptional regulation during globin switching. *P ≤ .05 as determined by Student t test comparing Tr4 heterozygous mutant with WT FACS-purified CD71+TER119+ cells.

Altered expression of select erythroid differentiation and proliferation genes upon reduction of TR4 activity in human and murine erythroid cells. (A,B) RNA-seq analysis from CD34+ cells infected with a sh-TR4 lentivirus, as previously reported,36  and RNA-seq results from control jumbled shRNA (shaded bars) or the shRNA-TR4 (white bars) values are represented as fragments per kilobase per million reads (FKPM).36  (C) qRT-PCR quantification of multiple genes expressed in erythroid cells of stage III (CD71+TER119+) cells sorted from both WT and Tr4+/− mouse total bone marrow. The fold changes were calculated using ∆∆Ct with WT set to 1 (indicated by the dashed line). (D) Schematic summarizing the known functions of TR4 in murine erythroid cells, including these data. Dashed lines indicate correlative data indicating Cdkn1c, Gata1, Alas2, Alad, and Gata2 are TR4 candidate target genes regulating erythroid proliferation and maturation. Solid arrows from TR4 to εy-globin and βh1-globin indicate direct transcriptional regulation during globin switching. *P ≤ .05 as determined by Student t test comparing Tr4 heterozygous mutant with WT FACS-purified CD71+TER119+ cells.

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