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. 2020 Sep 15;11:581011. doi: 10.3389/fphar.2020.581011

Figure 3.

Figure 3

HCK promotes the oligomerization of ASC. (A, B) Immunostaining of ASC in the LPS-primed PMs transfected with control siRNA (Mock) or siHCK for 3 h, followed by stimulation with nigericin for 30 min. (C) Immunoblotting analysis of the purified cross-linked ASC oligomers from the whole cell lysates in the LPS-primed (500 ng, 3 h) PMs transfected with control siRNA (Mock) or siHCK for 72 h, followed by stimulation with 5 μM nigericin for another 30 min (D, E) Immunostaining of ASC in the LPS-primed PMs after 1 h-incubation with A419259, followed by stimulation with Nigericin for 30 min. Nuclei were counterstained with DAPI. Fluorescence was imaged using confocal microscopy. (F) Immunoblotting analysis of the purified cross-linked ASC oligomers from the whole cell lysates in the LPS-primed (500 ng, 2 h) PMs treated with HCK inhibitor (A419259) for 1 h, followed by stimulation with 5 μM nigericin for another 30 min. Scale bar is 50 μm in all figures. Arrowheads denote ASC specks, and the percentage of ASC speck is shown in (B, E). Data from (A–F) are representative of at least three independent experiments. Data show means ± SEM, ***P < 0.001, Student’s t-test.