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. 2024 Apr 17;15:1294898. doi: 10.3389/fimmu.2024.1294898

Figure 2.

Figure 2

Interaction of HAdV-7 100 kDa protein with NLRP3. (A–D) HEK293T cells were co-transfected with 100 kDa-HA and NLRP3-Flag in (A), 100 kDa-HA and ASC in (B), and 100 kDa-HA and pro-caspase-1 in (C) and 100 kDa-HA and NLRP3-Flag in (D). Cell lysates were prepared and subjected to immunoprecipitation (IP) using EZview™ Red anti-HA beads (A–C) or anti-FLAG® M2 beads (D), followed by immunoblotting with anti-HA or anti-NLRP3, anti-ASC, and anti-caspase-1 antibody. (E) THP-1 macrophages were stably infected with CT-Lentivirus or 100 kDa-Lentivirus, differentiated into macrophages by PMA stimulation with for 48 (h) Cell lysates were immunoprecipitated using anti-FLAG® M2 beads and analyzed with anti-NLRP3 and anti-Flag antibody. (F) HEK293T cells were transfected with pCA7-NLRP3, GFP-100 kDa/GFP for 24 h. Subcellular localizations of NLRP3 (red), nucleus marker DAPI (blue), and GFP-100 kDa/GFP (green) was visualized using confocal microscopy. (G) PMA-differentiated THP-1 macrophages were stably infected with CT-lentivirus or 100 kDa-lentivirus. Subcellular localizations of NLRP3 (green), nucleus marker DAPI (blue), and 100 kDa-Flag/Flag (light blue) were visualized using confocal microscopy. Scale bar represents 10 μm.