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. 2022 Apr 18;298(6):101945. doi: 10.1016/j.jbc.2022.101945

Figure 1.

Figure 1

Phosphate-starvation results in massive induction of SLC20A1 protein abundance in HEK293T cells. Cells were phosphate starved (−Pi) for 48 h with phosphate-replete controls (+Pi). A, HEK293T cells were subjected to immunoblot analysis for SLC20A1 including siRNA-based knockdown for antibody validation (top lane) with β-actin loading control (middle lane). GAPDH siRNA was used as positive control for knockdown efficiency (bottom lane) and scrambled siRNA as negative control. Representative data of two experiments are shown. SLC20A1/β-actin abundance was determined by densitometry and normalized to the scrambled +Pi group (top). B, representative SLC20A1 immunofluorescence images (left) from sgControl-HEK293T (top) and sgSLC20A1-HEK293T cells (bottom) with DAPI as nuclear marker (center) and overlayed images on the right (SLC20A1 [green] and DAPI [blue]). The scale bars represent 10 μm. Representative data of three experiments are shown. C, SLC20A1 mRNA expression was normalized to 36B4. n = 6. Bars represent mean ± SD. Ct, average cycle threshold; DAPI, 4′,6-diamidino-2-phenylindole; HEK293T, human embryonic kidney 293T cell line.