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. 2021 Apr 1;9:624089. doi: 10.3389/fcell.2021.624089

FIGURE 4.

FIGURE 4

Luzp1–/– cells exhibit bigger nuclei and multiple centrioles. (A) Micrographs showing centrioles in wild-type Shh-LIGHT2 cells (WT), Shh-LIGHT2 cells lacking Luzp1 (Luzp1–/–) and Luzp1–/– cells rescued with Luzp1-YFP (+ LUZP1) analyzed during cycling conditions. Centrioles were visualized using gamma-tubulin antibody (gTub, magenta) and nuclei were counterstained using DAPI (blue). Scale bar, 5 μm. Cells containing 2 centrioles are marked with yellow arrowheads and those containing multiple centrioles with white asterisks. (B) Graphical representation of nuclear area of Shh-LIGHT2 WT (n = 178 micrographs; blue dots), Luzp1–/– (n = 185 micrographs; orange dots) and + LUZP1 cells (n = 90 micrographs; green dots). P-values were calculated using Kruskall-Wallis and Dunn’s multiple comparisons tests. (C) Graphical representation of the percentage of cells that exhibit more than 4 centrioles in (A). WT, n = 37 micrographs, blue dots; Luzp1–/–, n = 38 micrographs, orange dots; + LUZP1, n = 35 micrographs, green dots. P-values were calculated using Kruskall-Wallis and Dunn’s multiple comparisons tests. (D) Graphical representation of the percentage of cells that exhibit more than 4 centrioles in wild-type HEK 293FT (293WT) and HEK 293FT cells lacking Luzp1 (293LUZP1KO). 293WT, n = 9 micrographs, blue dots; 293LUZP1KO, n = 12 micrographs, orange dots. P-values were calculated using Mann Whitney-U test. The graphs in (B–D) represent the Mean and SEM. ∗P < 0.05; ∗∗∗P < 0.001; ****P < 0.0001.