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. 2016 Oct 31;291(52):26816–26836. doi: 10.1074/jbc.M116.719823

FIGURE 3.

FIGURE 3.

ANO6 is expressed abundantly in Caco-2 cells and mouse ileum. Quantitative PCR analysis of ANO1 and ANO6 in human colonic cells T84 and Caco-2 (A) and different parts of mouse intestine as indicated (B and C), which are shown as mRNA levels relative to levels of GAPDH and determined using the difference of PCR cycles to reach a threshold amplification (ΔCT), and the relative amount of the target mRNA are given as 2−ΔCT. CT, cycle threshold. Values are means ± S.E. of triplicate experiments. D, protein from total lysates of T84 and Caco-2 cell monolayers was resolved by 10% SDS-PAGE and immunoblotted with ANO6 antibody. ANO6 protein migrated in all cases with an apparent molecular mass of 106 kDa (top). Paraffin sections of mouse ileum were fixed and stained for ANO6 (green), and xy images were collected by confocal microscopy as described under “Experimental Procedures.” Image is representative of results for three experiments. Significant difference at p < 0.001 was compared between ANO1 and ANO6 (paired t test). NS, statistically not significant; ND, not determined.