Quantitative analysis confirmed that TGF-β1 alone did not significantly alter VCAM-1 expression compared with control (Dunnett’s test: mean difference = −1.12, 95% CI −3.85 to 1.61, p = 0.42), whereas TNF-α treatment resulted in a highly significant increase in VCAM-1 fluorescence intensity (mean difference = −22.24, 95% CI −24.57 to −19.90, p < 0.0001).
eached statistical significance compared with control in a one-way ANOVA followed by Dunnett’s test (adjusted p = 0.0002), while the combined TGF-β1 + TNF-α condition exhibited elevated nuclear pSMAD2/3 levels characterized by a broad high-intensity plateau on the fluorescence intensity profile but did not reach statistical significance, likely reflecting variability in nuclear size and signal distribution rather than absence of pathway activation ( Figure 2 ).