Figure 2. Macrophage VEGF-A expression is dependent on IL-1β expression during inflammatory conditions.
(A) BMDMs from control or myeloid IL-1β-deleted mice (mIL-1β KO) were primed with or without LPS (10 ng/mL) and exposed to cholesterol crystals (0 or 1000 μg/mL) for 24 h to induce inflammasome activation, followed by quantitative RT-PCR for relative IL-1β mRNA expression (***, p < 0.0001 compared to all others by ANOVA; n = 4 mice total, two males and two females).
(B) BMDMs treated as in (A), followed by ELISA on cell culture supernatants for secreted, mature IL-1β protein (***, p < 0.0001 compared to all others by ANOVA; n = 4 mice total, two males and two females).
(C) BMDMs treated as in (A), followed by quantitative RT-PCR for relative VEGF-A mRNA expression (***, p < 0.0001 compared to all others by ANOVA; n = 4 mice total, two males and two females).
(D) BMDMs treated as in (A), followed by ELISA on cell culture supernatants for secreted VEGF-A protein (***, p < 0.0001 compared to all others or between control and mIL-1β KO by ANOVA; n = 4 mice total, two males and two females).
(E) Primary mouse BMDMs from control or mIL-1β KO were treated with either LPS+IFN-γ or IL-4+IL-13 for 24 h, followed by ELISA on culture supernatants for secreted, mature IL-1β protein (***, p < 0.0001 compared to all others by ANOVA; n = 4 mice total, two males and two females).
(F) BMDMs treated as in (E) followed by quantitative RT-PCR for relative VEGF-A165a mRNA expression (***, p < 0.0001 compared to all others by ANOVA; n = 4 mice total, two males and two females).
(G) BMDMs treated as in (E) followed by quantitative RT-PCR for relative VEGF-A165b mRNA expression (**, p < 0.001 compared to all others by ANOVA; n = 4 mice total, two males and two females).
(H) BMDMs treated as in (E) followed by ELISA on culture supernatants for secreted VEGF-A protein (***, p < 0.0001 compared to all others by ANOVA; n = 4 mice total, two males and two females). Data, mean ± SD.