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. 2021 Jul 5;9:657406. doi: 10.3389/fcell.2021.657406

FIGURE 2.

FIGURE 2

Effect of cln5-deficiency on cell proliferation and viability in FM and FM-aa. (A) WT, cln5−, and cln5− + Cln5-GFP cells were placed in FM or FM-aa at a density of 1.0 × 105 cells/ml. Cell densities were measured every 24 h for 96 h. Data presented as mean density ± SEM (n ≥ 4). Statistical significance was assessed using two-way ANOVA followed by Bonferroni’s multiple comparisons test. **p-value < 0.01, ***p-value < 0.001, and ****p-value < 0.0001 at the indicated time points vs. WT. (B) Cells were grown in FM and FM-aa as described in panel (A). The CellTiter-Glo Luminescent Cell Viability Assay was used to quantify cell viability at the indicated time points. Statistical significance was determined using two-way ANOVA followed by Bonferroni’s multiple comparisons test. ***p-value < 0.001 and ****p-value < 0.0001 at the indicated time points vs. WT. (C) Cells (20 × 106 total) were harvested from HL5 suspension and placed in a 100 × 15 mm Petri dish containing HL5. After 1 h, the HL5 was removed and replaced with FM-aa. Images were taken after 24 h and are representative of four independent experiments. Scale bar = 50 μm.