Fig. 5: TGF-β mRNA-IAJD34-induced alterations in BAL cytokine production and cellular metabolic function.

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.