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. 2020 Aug 27;12(9):816. doi: 10.3390/pharmaceutics12090816

Figure 2.

Figure 2

IRBIT co-localized with NBCn1 and modulated the membrane expression of NBCn1. (A) Co-IP of NBCn1 and IRBIT in HEK293T cells. Cells were transfected with HA-tagged NBCn1 and GFP-tagged IRBIT, immunoprecipitated with GFP antibody, and blotted with NBCn1 antibody. β-Actin, input HA, and input GFP blots were used as loading controls (n = 3). (B) Immunostaining images of NBCn1 (red), GFP-IRBIT (green), and nucleus (DAPI, blue) in A549 cells. The scale bars represent 20 μm. (C) Magnified images of the dotted lines. (D) Analysis of NBCn1 intensity in the membrane expression. The bars indicate the mean ± SEM (n = 5). (E) Surface expression of NBCn1 in the presence of IRBIT in A549 cells. Input β-actin and input NBCn1 blots were used as loading controls. (F) The bars indicate the mean ± SEM of relative NBCn1 intensity normalized by input NBCn1 (n = 3).