milliplex max mouse cytokine/chemokine magnetic bead panel - premixed 32 plex - immunology multiplex assay (mcytmag-70kpx32)  (Millipore)

Journal: Nature Communications

Article Title: Targeted delivery of TGF-β mRNA to murine lung parenchyma using one-component ionizable amphiphilic Janus Dendrimers

doi: 10.1038/s41467-025-56448-y

Figure Lengend Snippet: Various doses of TGF-β mRNA-IAJD34 (10, 20, and 30 μg per mouse) were delivered to BALB/c mice, and BAL fluid was collected 24 h post-injection. A – D Cell- free BAL was evaluated for cytokines using a Milliplex Max Mouse Cytokine/Chemokine Magnetic Bead Panel - Premixed 32 Plex - Immunology Multiplex Assay. The concentration of ( A ) G-SCF, ( B ) IL-1α, ( C ) IL-9, and ( D ) TNFα are presented as pg/mL. E , F Cells from digested lung tissue were immunomagnetically separated based upon CD45 expression. CD45 + cells were isolated and analyzed metabolically using an Agilent Seahorse according to manufacturer’s guidelines; ( E ) ECAR was measured following injections of glucose, oligomycin (Oligo), and 2-DG, and glycolysis was determined following injection of glucose; ( F ) OCR was measured with injections of oligo, FCCP, and rotenone/antimycin (R/A) and normalized to non-mitochondrial energy production. Maximal respiration was determined following FCCP injection. Data are shown as mean ± SE, for all experiments n/group = 10/Naïve, 5/10 μg, 8/20 μg, 4/30ug. Data were evaluated for normality using a Shapiro- Wilk’s test and compared using a one-way ANOVA followed by Tukey’s multiple comparisons test. * indicates a significant difference from the control, p < 0.05. (( A ): Naïve vs 30 μg p = 0.0034; ( B ): Naïve vs 20 μg p = 0.0322, Naïve vs 30 μg p = 0.0003; ( C ): Naïve vs 20 μg p = 0.0129, Naïve vs 30 μg p = 0.0233; ( E ) Glycolysis: Naïve vs 20 μg p = 0.0001, Naïve vs 30 μg p < 0.0001). Black = Naïve, Red = 10 μg, Blue = 20 μg, Green = 30 μg. Source data are provided as a Source Data file.

Article Snippet: Cytokine in the first 1 mL of BAL fluid was performed by the University of Pennsylvania Human Immunology Core (RRID: SCR_022380) using a Milliplex Max Mouse Cytokine/Chemokine Magnetic Bead Panel - Premixed 32 Plex - Immunology Multiplex Assay (MCYTMAG-70K-PX32; MilliporeSigma, Burlington, MA).

Techniques: Injection, Multiplex Assay, Concentration Assay, Expressing, Isolation, Metabolic Labelling, Control

Journal: Nature Communications

Article Title: Targeted delivery of TGF-β mRNA to murine lung parenchyma using one-component ionizable amphiphilic Janus Dendrimers

doi: 10.1038/s41467-025-56448-y

Figure Lengend Snippet: BALB/c mice were exposed to PBS or bleomycin (ITB) and subsequently received either an empty IAJD34 or 10 µg TGF-β mRNA-IAJD34. Cell-free BAL fluid was collected 3 days post-exposure and injection. A – E Cell-free BAL fluid was evaluated for cytokines using a Milliplex Max Mouse Cytokine/Chemokine Magnetic Bead Panel - Premixed 32 Plex - Immunology Multiplex Assay. The concentrations of ( A ) G-SCF, ( B ) IL-6, ( C ) CXCL10, ( D ) IL-1α, and ( E ) IL-2 are presented as pg/mL. Date shown a mean ± SE, n/group = 7/PBS + Empty-IAJD34, 6/PBS + TGF-β mRNA + IAJD34, 10/ITB + Empty-IAJD34, 8/ITB + TGF-β mRNA+IAJD34. Data were evaluated for normality using a Shapiro Wilk’s test. If data were normally distributed, they were compared using a 2-way ANOVA followed by Šídák’s multiple comparisons test. Data points that were identified as 0 were set to the limit of detection according to manufacture values for statistical analysis. If data were not normally distributed, they were evaluated using a Kruskal Wallis test with Dunn’s multiple comparison test. * indicates a significant difference, p < 0.05. (( A ): PBS/Empty-IAJD34 vs ITB/Empty-IAJD34 p = 0.0231, ITB/Empty-IAJD34 vs ITB/TGF-β mRNA-IAJD34 p = 0.0300; ( B ): PBS/Empty-IAJD34 vs ITB/Empty-IAJD34 p = 0.0057, ITB/Empty-IAJD34 vs ITB/TGF-β mRNA-IAJD34 p = 0.0285; (C): PBS/Empty-IAJD34 vs ITB/Empty-IAJD34 p = 0.0168; ( D ): PBS/Empty-IAJD34 vs ITB/Empty-IAJD34 p = 0.0296; ( E ) PBS vs ITB p = 0.0107). Source data are provided as a Source Data file.

Article Snippet: Cytokine in the first 1 mL of BAL fluid was performed by the University of Pennsylvania Human Immunology Core (RRID: SCR_022380) using a Milliplex Max Mouse Cytokine/Chemokine Magnetic Bead Panel - Premixed 32 Plex - Immunology Multiplex Assay (MCYTMAG-70K-PX32; MilliporeSigma, Burlington, MA).

Techniques: Injection, Multiplex Assay, Comparison