FIGURE 5.
Knockdown of endogenous annexin A6 causes increased Ca2+ entry in HEK293 cells. A, HEK293 cells were co-transfected with shRNA plasmids expressing annexin A6-specific siRNA and mCherry. Expression levels of endogenous annexin A6 were examined by immunofluorescence 48 h post transfection. mCherry-positive cells do not express annexin A6 (arrows). B, HEK293 cells co-expressing annexin A6 shRNA together with mCherry or scrambled shRNA control together with mCherry-tH were co-cultured, loaded with Fluo-3/AM, and stimulated with 1 μm TG. SOCE was activated by adding 0.5, 2, 5, 7, and 10 mm [Ca2+]ex. In a representative experiment, responses in mCherry-expressing annexin A6-negative cells (gray line) were compared with control mCherry-tH cells (black line). C, an average of four independent experiments (n = 14 recordings) is shown in the graph.