Fig. 10: MZT2B knockdown impairs NSCLC xenograft growth in vivo.

Analysis of subcutaneous xenografts derived from primary pNSCLC1-1 cells stably expressing control shRNA (shC) or shRNA targeting MZT2B (kdMZT2B-sh2) engrafted into nude mice. Longitudinal monitoring of tumor volume over 42 days post-initiation (Day-0 defined as three weeks post-engraftment) (A). Calculation of the derived daily tumor growth (B). Assessment of explanted tumor mass at Day-42 (C). Longitudinal monitoring of animal body weights (D). Validation of target modulation within tumor tissues explanted at Day-24 and Day-30 encompassed qRT-PCR analysis of MZT2B and MZT2A mRNA levels (E) and immunoblotting analysis with quantification of MZT2B and MZT2A protein levels (F and G). Further analyses of tumor characteristics included IHC assessment of Ki-67-positive proliferating cells in tumor sections (H), quantification of cytosolic Cytochrome c levels in tumor lysates (I), immunoblotting analysis with quantification of cleaved Caspase-3 and cleaved PARP1 protein levels in tumor lysates (J), and TUNEL assay for detection of apoptotic cells in tumor sections (K). The COX5B mRNA and COX5B protein levels (L, M) were also analyzed. Analyses of mitochondrial function and redox state performed on lysates from tumors explanted at Day-24 and Day-30 encompassed measurement of total cellular ATP contents (N), determination of the cellular GSH/GSSG ratio (O), and measurement of ssDNA intensity (P). Error bars represent the mean ± standard deviation (SD), with statistical significance (*P < 0.05 when comparing to “shC” group). “n.s.” stands for P > 0.05. Data presented in (A–D) were derived from n = 10 mice per group. For (E–P), analyses were conducted on five distinct tissues randomly selected from each xenograft (n = 5). Scale bar = 100 μm.