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. 2020 Oct 1;34(19-20):1345–1358. doi: 10.1101/gad.340661.120

Figure 6.

Figure 6.

Uptake of necrotic cells by macropinocytosis is dependent on the Yap/Taz transcriptional target Axl. (A) Immunoblot of whole cell lysates from WT KRPC or Yap/Taz dKO cells transduced with retroviruses containing a dox-inducible Yap or Taz cDNA after culture for 24 h in 100 ng/mL doxycycline. (B) Histogram showing engulfment of cypHer5E-stained necrotic FL5.12 cells by KRPCS cells or parental KRPC after 2 h in complete medium, as measured by flow cytometry. (C) Bar plot represents mean ± SD frequency of cypHer5E-positive cells from three technical replicates. (***) P < 0.0001, unpaired two-tailed Student's t-test. (D) Engulfment of cypHer5E-stained necrotic FL5.12 cells after 6 h by WT or Yap/Taz dKO KRPC cells. Cells were transduced with retroviruses containing a doxycycline-inducible Yap or Taz cDNA and necrotic cell engulfment was measured after 6 h in the presence of 100 nM LDC1267 or vehicle. Assay was performed following 18 h culture in 100 ng/mL doxycycline. Bars represent mean ± SD cypHer5E-positive cells, as measured by flow cytometry, from three technical replicates. (E) Engulfment of cypHer5E-stained necrotic FL5.12 cells after 6 h by WT KRPC or Yap/Taz dKO cells transduced with retroviruses containing a doxycycline-inducible mouse Axl cDNA (either isoform 1 or 3). Assay was performed following an 18-h culture in 100 ng/mL doxycycline. Bars represent mean ± SD cypHer5E-positive cells, as measured by flow cytometry, from three technical replicates. (***) P < 0.0001, one-way ANOVA.