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. 2021 Sep 8;22(11):e53732. doi: 10.15252/embr.202153732

Figure 7. Generation of Nrl.Cre+/−/TdTomatofloxed aggregate chimeras reveals intercellular exchange of Cre between photoreceptors in vivo .

Figure 7

  • A
    Schematic representation of Nrl.Cre+/−/TdTomato+/− floxed chimera generation via morula aggregation, which results in chimeric mosaicism in the neural retina. Recombination and expression of TdTomato is only possible if Nrl.Cre+/− photoreceptors exchange material (Cre) with TdTomatofloxed cells via intercellular communication.
  • B
    Representative tile scan of uninjected Nrl.Cre+/− /TdTomato+/− chimera showing endogenous recombination (white); ROIs, as indicated with dashed or dot boxes; Scale bars = 50 µm.
  • C, D
    ROI from uninjected Nrl.Cre+/− /TdTomato+/− chimera showing spontaneous recombination presenting in mosaic stripe patterning; Scale bar = 50 µm in (C) and 10 µm in (D).
  • E, F
    Representative images of Nrl.Cre+/− /TdTomato+/− chimera after receiving a subretinal injection of AAV‐Nrl.Cre. The injected eyes show widespread recombination, which also shows the anticipated mosaic stripe patterning; Scale bar = 50 µm in (E) and 10 µm in (F).
  • G
    Quantification of number of recombined cells in uninjected (29,825 cells ± 2,031) and AAV‐Nrl.Cre‐injected (131,562 ± 8,478) Nrl‐Cre+/− /TdTomato+/− chimeras, compared with uninjected Nrl‐Cre −/− (wt)/TdTomato+/− chimera (0 ± 0). Graph shows mean ± SD.

Data information: white = TdTomato+ve recombined cells; blue = nuclei. RPE‐ Retinal Pigment Epithelium; ONL—outer nuclear layer; INL—inner nuclear layer; GCL—Ganglion Cell Layer.