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. Author manuscript; available in PMC: 2022 Mar 15.
Published in final edited form as: J Immunol. 2021 Feb 10;206(6):1329–1336. doi: 10.4049/jimmunol.2000969

Figure 3. Tyrosine 146 of ASC CARD is critical for IL-1β processing.

Figure 3.

THP-1 cells were engineered using CRISPR/Cas9 technology to create ASC KO THP-1 cells. These KO cells were then transduced with a lentivirus knock-in (KI) of either wild type ASC isoform “a” (ASC KI) or the Y146A mutant of ASC “a” (ASC Y146A). The relative expression of ASC in these modified THP-1 cells is shown by ASC immunoblot and the expression of proIL-1β after priming with LPS by immunoblot for IL-1β (A). IL-1β release from THP-1 cells stimulated with LPS+ATP for 3h+30 min is shown in (B) and after LPS overnight in (C) was detected by ELISA. LDH and IL-18 release from cells stimulated with LPS for 30 min followed by ATP or nigericin for another 30 min shown in (D, E and F), respectively. n=5–7 independent experiments. *** p<0.001, ** p<0.01, * p<0.05 as compared to Cas9 control cells.