The figure contains multiple scatter bar plots. Panel A shows IFN beta release in response to Poly I:C treatment, with the y-axis labeled as IFN beta release (ng/mL) and the x-axis indicating different conditions (DMEM and hypo-osmotic media) for WT and VRAC KO BMDMs. Panel B shows IFN beta release in response to LPS treatment, with similar axes and conditions. Panel C shows IFN beta release in response to Imiquimod (IQ) treatment, and Panel D shows IFN beta release in response to CpG-DNA treatment, both with similar axes and conditions. Panels E to H show IL-6 release in response to the same treatments and conditions as Panels A to D. Panel I shows LDH release under various conditions and treatments, with the y-axis labeled as LDH release (percent whole well lysis). Panels J to M show IL-6 release in WT BMDMs under different osmotic conditions (DMEM, hypo-osmotic, isotonic, and hyper-osmotic media) and treatments (Poly I:C, LPS, IQ, CpG-DNA). Panels N to O show IFN beta release in WT BMDMs under the same osmotic conditions and treatments as Panels J to M. The graphs indicate significant differences in cytokine and LDH release under different conditions and treatments, with statistical significance marked by asterisks.
Cell volume regulates type I interferon production in response to diverse pathogen-mediated molecular patterns. (A–D) IFNβ release in the supernatant of WT and VRAC KO BMDMs incubated in DMEM or hypo-osmotic media (50% vol/vol H2O in DMEM) and treated with poly I:C (5 μg ml−1) (A), LPS (1 μg ml−1) (B), imiquimod (IQ, 25 µM) (C), or CpG-DNA (CpG, 5 μg ml−1) (D) for 6 h (n = 9 for poly I:C, LPS, CpG; n = 4 for IQ). (E–H) IL-6 release in the supernatant of WT and VRAC KO BMDMs incubated in DMEM or hypo-osmotic media (50% vol/vol H2O in DMEM) and treated with poly I:C (5 μg ml−1) (E), LPS (1 μg ml−1) (F), imiquimod (25 µM) (G), or CpG-DNA (5 μg ml−1) (H) for 6 h (n = 9 for poly I:C, LPS, CpG; n = 4 for IQ). (I) LDH release from WT and VRAC KO BMDMs incubated in iso-osmotic or hypo-osmotic media in the presence of poly I:C (I:C, 5 µg ml−1), LPS (1 µg ml−1), IQ (IQ, 25 µM), or CpG-DNA (CpG, 5 µg ml−1) for 6 h (n = 6). (J–M) IL-6 release in the supernatant of WT BMDMs incubated in either DMEM, hypo-osmotic media (50% vol/vol H2O in DMEM), isotonic media (50% vol/vol 150 mM NaCl), or hyperosmotic media (50% vol/vol 300 mM NaCl) and treated with poly I:C (5 μg ml−1) (J), LPS (1 μg ml−1) (K), imiquimod (IQ, 25 µM) (L), or CpG-DNA (CpG, 5 μg ml−1) (M) for 6 h (n = 6). (N and O) IFNβ release in the supernatant of WT BMDMs incubated in either DMEM, hypo-osmotic media (50% vol/vol H2O in DMEM), isotonic media (50% vol/vol 150 mM NaCl), or hyperosmotic media (50% vol/vol 300 mM NaCl) and treated with poly I:C (5 μg ml−1) (N) or LPS (1 μg ml−1) (O) for 6 h (n = 6). *P < 0.05, **P < 0.01, ***P < 0.001, determined by a two-way ANOVA with Sidak’s post hoc analysis (7A–7H) or Tukey’s post hoc analysis (7I), or by a one-way ANOVA with Tukey’s post hoc analysis (7J–7O). Values shown are the mean ± the SEM.