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. 2019 Sep 17;8:e48003. doi: 10.7554/eLife.48003

Figure 6. Knockdown and inhibition of TRPV6 and PP2A increases human colon carcinoma cell proliferation.

(A) LoVo cells transfected with scrambled siRNA or TRPV6 targeting siRNA were synchronized by serum starvation followed by serum re-stimulation. Cells were analyzed by flow cytometry analysis after propidium iodide staining. Percentage of S-phase cells are calculated and shown. Mean ± SEM, n = 3. **, p<0.01 by unpaired t-test. (B) LoVo cells were synchronized by serum starvation. They were re-stimulated with 2% FBS medium containing Ruthenium Red (RR, 100 µM), GdCl3 (100 µM), BAPTA-AM (100 µM), Okadaic acid (OA, 20 nM) or DMSO for 48 hr and analyzed by flow cytometry analysis after propidium iodide staining. Percentage of S-phase cells are shown. Mean ± SEM, n = 3. (C) LoVo cells transfected with the indicated DN-PP2A constructs were synchronized by serum starvation followed by serum re-stimulation. Cells were analyzed by flow cytometry analysis after propidium iodide staining. Percentage of S-phase cells are calculated and shown. Mean ± SEM, n = 3. ****, ***, ** indicate P<0.0001, P<0.001, P<0.01 by one-way ANOVA followed by Tukey’s multiple comparison test in (B) and (C). (D) Schematic diagram of the proposed model. See text for details.

Figure 6—source data 1. Excel spreadsheet containing quantitative data for Figure 6.

Figure 6.

Figure 6—figure supplement 1. Cell cycle analysis profiles for experiments shown in Figure 6A.

Figure 6—figure supplement 1.

The PI fluorescence intensity is given in arbitrary units (a.u.) on the X-axis, and the Y-axis indicates cell counts normalized to percent of the max as measured by flow cytometry.
Figure 6—figure supplement 2. Cell cycle analysis profiles for inhibitor experiments shown in Figure 6B.

Figure 6—figure supplement 2.

Figure 6—figure supplement 3. LoVo cell viability.

Figure 6—figure supplement 3.

LoVo cells were synchronized by serum starvation. They were re-stimulated with 2% FBS medium containing Ruthenium Red (RR, 100 µM), GdCl3 (100 µM), BAPTA-AM (100 µM), Okadaic acid (OA, 20 nM) or DMSO for 48 hr and analyzed by MTT assays. Mean ± SEM, n = 3.
Figure 6—figure supplement 4. Cell cycle analysis profiles for experiments shown in Figure 6C.

Figure 6—figure supplement 4.