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. 2021 Jul 1;11:671431. doi: 10.3389/fonc.2021.671431

Figure 6.

Figure 6

USP9X depletion destabilizes centrosomal proteins, and induces pericentrin-rich foci amplification in response to relatively high-LET protons. (A, C, D) HeLa or (B, E, F) UMSCC74A cells were treated with USP9X or a non-targeting (NT) control siRNA for 48 h, and unirradiated designated (C) or irradiated with 4 Gy relatively high-LET or low-LET protons and allowed to repair for the time points indicated post-irradiation. (A, B) Whole cell extracts were analyzed by immunoblotting using the indicated antibodies. Red and black text refers to time post-irradiation following relatively high-LET and low-LET protons, respectively. Protein levels of CEP131 and CEP55 relative to tubulin and normalized to the unirradiated NT siRNA control treated cells which was set to 1.0 are shown. (C, E) Analysis of pericentrin was performed by immunofluorescence 48 h post-irradiation. Shown is the mean percentage of cells with >2 pericentrin-rich foci/nucleus ± S.D. *p < 0.03 as analyzed by a two-sample t-test. (D, F) Representative images of pericentrin staining (green stain) and nuclei (blue stain; DAPI).