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. 2020 Dec 22;40(4):e104729. doi: 10.15252/embj.2020104729

Figure 7. Bptf and Tbx3 alternative exon loss can rescue defective differentiation in hnRNPLL‐KO ES cells.

Figure 7

  1. RT–PCR validation of Bptf and Tbx3 alternative exon deletion, separately in hnRNPLL‐KO H8 cells (upper) and simultaneously in hnRNPLL‐KO H8 cells (lower).
  2. AP staining of H8‐Bptf or H8‐Tbx3 exon‐KO ES cells after LIF withdrawal. hnRNPLL‐H8, hnRNPLL‐KO ES cells; H8‐Bptf KO, alternative Bptf exon was deleted in hnRNPLL‐KO H8 cells; H8‐Tbx3 KO, alternative Tbx3 exon was deleted in hnRNPLL‐KO H8 cells; H8‐Bptf/Tbx3 KO, alternative Bptf or Tbx3 exons were simultaneously deleted in hnRNPLL‐KO H8 cells. Scale bar, 200 μm.
  3. Colony formation assay of H8‐Bptf or H8‐Tbx3 exon‐KO ES cells after LIF withdrawal. The data represent the means ± SD (n = 3, from biological repeats).
  4. Hematoxylin and eosin labeling of teratomas derived from WT and H8‐Bptf or H8‐Tbx3 exon‐KO ES cells. The blue triangle represent ectoderm, the dark one represent mesoderm, and the green one represent endoderm. Scale bar, 100 μm.
  5. Oct4 immunohistochemical staining of teratomas derived from WT and H8‐Bptf or H8‐Tbx3 exon‐KO ES cells. Representative Oct4‐positive regions are shown (left). Scale bar, 100 µm. The percentage Oct4‐positive region (%) is shown (right). The data represent the means ± SD (n = 3 biological repeats). **P < 0.01; ns, not significant. t‐Test was used to determine the significance.
  6. Schematic representation of hnRNPLL regulating the AS of crucial genes that have an important role in mediating exit from pluripotency.