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

Fig. 4. Pharmacological inhibition of STK39/OXSR1 increases NLRP3 inflammasome activation in macrophages.

Fig. 4

Propidium iodide uptake of LPS-primed (4 h) primary wild type BMDMs treated with or without 1 uL of DMSO or 2 µM Closantel or Rafoxanide for 10 m prior to treatment with ATP (a) or imiquimod (b). P values for a are 0.011432 and 0.00492; b are 0.00162 and 0.00091. IL-1β (c) and LDH (d) release in culture supernatants of LPS-primed (4 h) primary wild type bone marrow macrophages treated with or without 1 uL of DMSO or 2 µM Closantel or Rafoxanide for 10 m prior to treatment with ATP or imiquimod. P values for c are 0.00291, 0.00585, 0.00707, and 0.0048; d are 0.00042, 0.00129, 0.00162, and 0.00071. Propidium iodide uptake of LPS-primed (15 m) immortalized wild type BMDMs treated with or without 1 uL of DMSO or 2 µM Closantel or Rafoxanide for 10 m prior to treatment with ATP (e) or imiquimod (f). P values for e are 0.00126 and 0.00149; f are 0.00164 and 0.000296. g Immunoblots of caspase-1 p20 released in culture supernatants (Sup) or procaspase-1 and NLRP3 in cell lysates (Lys) of LPS-primed (15 m) treated with or without 1 uL of DMSO or 2 µM Closantel or Rafoxanide for 10 m prior to treatment with ATP or nigericin as indicated. The split caspase 1 p20 represents two exposures of the same membrane. Results are representative of at least three independent experiments performed in duplicate or triplicate. Error bars in a–f 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.