Fig. 2: Evaluation of dose-dependent toxicity of TGFβ mRNA delivery with IAJD34.

Various doses of TGFβ mRNA-IAJD34 (10, 20, and 30 μg per mouse) were delivered to BALB/c mice and compared to naïve animals. At 24 h post-injection, BAL fluid and cells, whole lung tissue, and serum were collected. A BAL cells were fixed and stained using a Hema 3 Stat kit. Total BAL protein, differential BAL cell count, and BAL phospholipid levels were quantified as markers of lung injury: MP, Macrophage, NP, neutrophil, LP, Lymphocyte. B Representative cell differentials are shown. C Formaldehyde-fixed lung tissue was processed, stained with H&E, and scored by a board-certified pathologist. Representative H&E images are shown. D IL-6 was measured in BAL fluid via ELISA according to manufacturer’s guidelines (n/group = 10/Naïve, 5/10 μg, 7/20 μg, 3/30ug). E ALT and AST were measured in serum via ELSIA according to the manufacturer’s guidelines (n/group = 5/Naïve, 5/20 μg, 5/30 μg). Data are presented as mean ± SE, n = 3-10/group from 2 independent experiments. For (A, D, and E), samples were compared using the Shapiro-Wilk test for normality followed by an ordinary one-way ANOVA with Tukey’s Multiple comparisons where *indicates a significant difference from the naïve control, p < 0.05. ((A) BAL protein: Naïve vs 30 μg p = 0.0001; NP: Naïve vs 30 μg p = 0.0134; (E) ALT: Naïve vs 30 μg p = 0.0164). Scale bar = 50 µm. Source data are provided as a Source Data file.