Figure 7.
eTNS1 is required to assemble F-actin into the enucleosome during enucleation of human erythroblasts. (A) Maximum intensity projections of Airyscan Z-stacks of polarized and enucleating human erythroblasts from day-14 cultures stained for F-actin (phalloidin; magenta), eTNS1 (red), GPA (green), and nuclei (Hoechst; blue). F-actin in eTNS1− cells assembled into mislocalized foci (yellow arrow), cables (red arrow), or was absent. Scale bar, 5 μm. (B) Quantification of F-actin intensity in eTNS1− and eTNS1+ erythroblasts from eTNS1-KO cultures at days 14 and 17 using the Zeiss CD7. (C) Quantification of F-actin intensity normalized to GPA intensity of eTNS1+ and eTNS1− erythroblasts from eTNS1-KO cultures at days 14 and 17. (B-C) Plots reflect the mean ± SD of 1500 eTNS1+ and eTNS1− erythroblasts from 3 independent eTNS1 CRISPR KO experiments. ∗∗∗∗P < .0001. a.u., arbitrary units.

eTNS1 is required to assemble F-actin into the enucleosome during enucleation of human erythroblasts. (A) Maximum intensity projections of Airyscan Z-stacks of polarized and enucleating human erythroblasts from day-14 cultures stained for F-actin (phalloidin; magenta), eTNS1 (red), GPA (green), and nuclei (Hoechst; blue). F-actin in eTNS1− cells assembled into mislocalized foci (yellow arrow), cables (red arrow), or was absent. Scale bar, 5 μm. (B) Quantification of F-actin intensity in eTNS1− and eTNS1+ erythroblasts from eTNS1-KO cultures at days 14 and 17 using the Zeiss CD7. (C) Quantification of F-actin intensity normalized to GPA intensity of eTNS1+ and eTNS1− erythroblasts from eTNS1-KO cultures at days 14 and 17. (B-C) Plots reflect the mean ± SD of 1500 eTNS1+ and eTNS1− erythroblasts from 3 independent eTNS1 CRISPR KO experiments. ∗∗∗∗P < .0001. a.u., arbitrary units.

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