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. 2020 Sep 4;68(10):679–690. doi: 10.1369/0022155420955246

Figure 4.

Figure 4.

PLA for myc-p11 binding to GFP-TASK channels. (A) Confocal images of GFP-TASK1 or GFP-TASK3 and PLA reaction between GFP and myc in PC12 cells exogenously expressing myc-p11 and GFP-TASK1 (T1G) or GFP-TASK3 (T3G). (B) Summaries of PLA products in the total area and at the cell periphery of PC12 cells. Data are mean ± SEM; Mann–Whitney rank-sum test (T1G: n=10 from three culture dishes; T3G: n=10 from two culture dishes). Bars are 5 µm. (C) Exogenous expression of GFP-TASK1 or GFP-TASK3 in PC12 cells. First and second columns represent confocal images of GFP and TASK1- or TASK3-like IR material, respectively; third represents merge of first and second images. GFP-TASK and TASK-like IR material were visible as GFP and rhodamine-like fluorescence, respectively. (D) TASK1- and TASK3-like IR material are plotted against those of GFP fluorescence (TASK1: n=20 from five culture dishes; TASK3: n=19 from four culture dishes). Linear regression lines are y = −7911 + 2.811x (correlation coefficient r = 0.996) and y = −13330 + 2.662x (r = 0.994) for TASK1 and TASK3, respectively. (E) Immunocytochemical staining for TASK1 and TASK3 endogenously expressed in PC12 cells. First and second columns represent confocal images and merge of fluorescence and DIC images. (F) TASK1- and TASK3-like IR material in PC12 cells. Data are mean ± SEM (n=20 from two culture dishes for each of TASK1 and TASK3). Bars are 5 µm. Abbreviations: DIC, differential interference contrast; GFP, green fluorescent protein; IR, immunoreactive; PC, pheochromocytoma; PLA, proximity ligation assay; TASK, TWIK-related acid-sensitive K+.