Skip to main content
. Author manuscript; available in PMC: 2019 Feb 15.
Published in final edited form as: Mol Cell. 2018 Feb 15;69(4):622–635.e6. doi: 10.1016/j.molcel.2018.01.011

Figure 1. DKO of TIA1 and TIAL1 in HEK293 cells abrogates cell cycle progression and causes apoptotic cell death.

Figure 1

A) Domain organization of human TIA1 family proteins. Protein length in amino acids (aa) is indicated. RRMs, RNA recognition motif; PrLD, prion-like domain. B) Verification of TIA1/TIAL1 DKO in HEK293 cells expressing either FH-tagged TIAL1 or TIA1 cultured with or without Dox. TUBB: Immunoblot (IB) for Tubulin beta chain. C) DKO/FH-TIAL1 and DKO/FH-TIA1 cells cultured without Dox (−Dox) over a 10-day time-course. D) Quantification of FH-tagged protein levels in DKO/FH-TIAL1 and DKO/FH-TIA1 cells upon Dox depletion based on IB (n=3) as shown in (B). E) Cell cycle analyses of DKO/FH-TIAL1 or DKO/FH-TIA1 cells cultured without Dox for up to 8 days. F) IB for phosphorylated H2B, cleaved CASP3 and cleaved PARP on DKO/FH-TIAL1 cell lysates cultured without Dox for up to 8 days. G) Fluorescence microscopy images of unsynchronized DKO/FH-TIAL1 cells cultured with (+Dox) or without Dox for 6 days (−Dox). Nuclei stained with DAPI. Scale bar, 2 μm. H) IF on TUBB to visualize centrosome formation in DKO/FH-TIAL1 cells. DAPI was used to stain nuclear DNA. Right panels depict colored overlays (DNA, blue; TUBB, green) of single grey scale images. Scale bar, 2 μm. I) For DKO/FH-TIAL1 cells cultured with (0 days) or without Dox (8 days), 100 mitotic cells were scored and the percentages of cells showing 1 (monopolar), 2 (bipolar asymmetric or symmetric), 3 (tripolar), or 4 (quadrupolar) centrosomes were calculated.