Fig 2: ABCC5, CLDN1, and CSTA expression in LUSC and paracancerous tissues. (a) mRNA expression levels of ABCC5, CLDN1, and CSTA in LUSC and paracancerous tissues. (b) Protein expression levels of ABCC5, CLDN1, and CSTA in LUSC and paracancerous tissues. (c) Representative images of immunohistochemistry staining for ABCC5, CLDN1, and CSTA in LUSC and paracancerous tissues. Magnification, ×200. Data are displayed as the mean ± standard deviation. Student's t-test was performed.∗P < 0.05 and ∗∗∗P < 0.001. LUSC, lung squamous cell carcinoma; CLDN1, Claudin 1; ABCC5, ATP-binding cassette subfamily C member 5; CSTA, Cystatin A.

Fig 3: Validation of key host gene expression in 3D vs. 2D cultures. (A) RT-qPCR validation of top DEGs and HPV45 genes in HPV45+ primary cervical cancer cells (MCW-3) grown in 3D vs. 2D culture. Results shown as mean fold change in 3D relative to 2D (error bars: SE, n = 3 biological x 3 technical replicates). p-value < 0.01*, <0.0001***, <0.00001 ****; (B) Western blot analysis of HPV-independent genes, CASP14, S100A9, ALPP, CLDN1, and IL18, with differential expression in 2D-3D cultures.