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Dose-dependent inflammasome activation and cytotoxicity in LPS-primed mouse macrophages and dendritic cells stimulated with AH or AP. BMDMs ( a–c ) and BMDCs ( d – f ) derived <t>from</t> <t>CD-1</t> mice were incubated in medium only (M) or primed with LPS and stimulated with increasing concentrations of AH or AP for 24 h. ( a , d ). Cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) release. ( b , e ) IL-1β secretion was quantified by ELISA. ( c , f ) Western blot analysis for IL-1β of supernatants (Sup) and lysates (Lys). Ten µg of total protein was loaded per lane for BMDMs (c) and 20 µg for BMDCs (f). Bars represent the mean ± SEM of triplicate wells. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).
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Dose-dependent inflammasome activation and cytotoxicity in LPS-primed mouse macrophages and dendritic cells stimulated with AH or AP. BMDMs ( a–c ) and BMDCs ( d – f ) derived <t>from</t> <t>CD-1</t> mice were incubated in medium only (M) or primed with LPS and stimulated with increasing concentrations of AH or AP for 24 h. ( a , d ). Cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) release. ( b , e ) IL-1β secretion was quantified by ELISA. ( c , f ) Western blot analysis for IL-1β of supernatants (Sup) and lysates (Lys). Ten µg of total protein was loaded per lane for BMDMs (c) and 20 µg for BMDCs (f). Bars represent the mean ± SEM of triplicate wells. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).
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Dose-dependent inflammasome activation and cytotoxicity in LPS-primed mouse macrophages and dendritic cells stimulated with AH or AP. BMDMs ( a–c ) and BMDCs ( d – f ) derived from CD-1 mice were incubated in medium only (M) or primed with LPS and stimulated with increasing concentrations of AH or AP for 24 h. ( a , d ). Cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) release. ( b , e ) IL-1β secretion was quantified by ELISA. ( c , f ) Western blot analysis for IL-1β of supernatants (Sup) and lysates (Lys). Ten µg of total protein was loaded per lane for BMDMs (c) and 20 µg for BMDCs (f). Bars represent the mean ± SEM of triplicate wells. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

Journal: Scientific Reports

Article Title: Aluminum adjuvants differentially induce IL-1β release in vitro yet share NLRP3 inflammasome-independent adjuvant effects in vivo

doi: 10.1038/s41598-025-34660-6

Figure Lengend Snippet: Dose-dependent inflammasome activation and cytotoxicity in LPS-primed mouse macrophages and dendritic cells stimulated with AH or AP. BMDMs ( a–c ) and BMDCs ( d – f ) derived from CD-1 mice were incubated in medium only (M) or primed with LPS and stimulated with increasing concentrations of AH or AP for 24 h. ( a , d ). Cytotoxicity was assessed by measuring lactate dehydrogenase (LDH) release. ( b , e ) IL-1β secretion was quantified by ELISA. ( c , f ) Western blot analysis for IL-1β of supernatants (Sup) and lysates (Lys). Ten µg of total protein was loaded per lane for BMDMs (c) and 20 µg for BMDCs (f). Bars represent the mean ± SEM of triplicate wells. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****).

Article Snippet: Female Hsd:ICR (CD-1®) (CD-1) mice were purchased from Inotiv (Indianapolis, IN).

Techniques: Activation Assay, Derivative Assay, Incubation, Enzyme-linked Immunosorbent Assay, Western Blot

The antibody response to protein antigens injected with aluminum adjuvants does not require functional NLRP3. ( A , B ) C57BL/6J WT and Nlrp3 -/- mice were injected twice with OVA and AH ( A ) or nucleoprotein of SARS-CoV-2 (NP) and AP ( B ). The graphs show the mean ± SE of four mice per group. ( C , D ) CD-1 mice were treated with MCC950 or untreated and then injected twice with OVA and AH ( C ) or nucleoprotein of SARS-CoV-2 (NP) and AP ( D ). The graphs show data from individual mice and the mean ± SE of 4–5 mice per group. ns – not significant (Student’s t-test, p > 0.05).

Journal: Scientific Reports

Article Title: Aluminum adjuvants differentially induce IL-1β release in vitro yet share NLRP3 inflammasome-independent adjuvant effects in vivo

doi: 10.1038/s41598-025-34660-6

Figure Lengend Snippet: The antibody response to protein antigens injected with aluminum adjuvants does not require functional NLRP3. ( A , B ) C57BL/6J WT and Nlrp3 -/- mice were injected twice with OVA and AH ( A ) or nucleoprotein of SARS-CoV-2 (NP) and AP ( B ). The graphs show the mean ± SE of four mice per group. ( C , D ) CD-1 mice were treated with MCC950 or untreated and then injected twice with OVA and AH ( C ) or nucleoprotein of SARS-CoV-2 (NP) and AP ( D ). The graphs show data from individual mice and the mean ± SE of 4–5 mice per group. ns – not significant (Student’s t-test, p > 0.05).

Article Snippet: Female Hsd:ICR (CD-1®) (CD-1) mice were purchased from Inotiv (Indianapolis, IN).

Techniques: Injection, Functional Assay