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. 2021 Jul 27;12:4546. doi: 10.1038/s41467-021-24784-4

Fig. 2. Tamoxifen-induced WNK1 knockout increases NLRP3 inflammasome activation in macrophages.

Fig. 2

a Immunoblots of caspase-1 in culture supernatant (Sup) and cell lysates (Lys) of immortalized Wnk1flox/flox-CreERT2+ (WNK1 KO) or Wnk1+/+ (WT) macrophages treated with tamoxifen for 48 h prior to stimulation with Pam3CSK4, Poly(I:C) or LPS for the indicated times followed by ATP (45 min). LDH release of immortalized Wnk1flox/flox-CreERT2+ (WNK1 KO) or Wnk1+/+ (WT) macrophages treated with tamoxifen for 48 h prior to stimulation with Pam3CSK4 (b) or Poly(I:C) (c) for the indicated times followed by ATP (45 min). P values for b are 0.321, 0.00473, 0.0036, 0.0475, 0.0754, and 0.0027; c are 0.716, 0.0055, 0.0004, 0.0006, 0.0001, and 0.0191. d Immunoblots of caspase-1 in culture supernatant (Sup) and cell lysates (Lys) of immortalized Wnk1flox/flox-CreERT2+ (WNK1 KO) or Wnk1+/+ (WT) macrophages treated with tamoxifen for 48 h prior to stimulation with LPS for 3 h followed by MSU (40 uL or 80 uL 5 mg mL− MSU) for 80 m. IL-1β release (e) and propidium iodide uptake (f, g) of immortalized Wnk1flox/flox-CreERT2+ (WNK1 KO) or Wnk1+/+ (WT) macrophages treated with tamoxifen for 48 h prior to stimulation with LPS for 3 h followed by MSU (40 uL (f) or 80 uL (g) 5 mg mL− MSU) for 80 m. P values for e are 0.061, 0.00077, and 0.00094; f is 0.0005; g is 0.00034. Results are representative of at least three independent experiments performed in duplicate or triplicate. Error bars in b, c, e–g are presented as mean values ± standard deviation (S.D.), with n = 3. Two-sided Student’s t test, *p < 0.05, **p < 0.005, ***p < 0.0005.