Fig. 1

The efficacy of Pulsatilla saponin D (PSD) in inhibiting the proliferation of paclitaxel-resistant cells. (A): A549-PR, A549, NCI-H1299 and NCI-H1299-PR cells were subjected to different concentrations of PSD (0, 1, 5, 10, 20 µM), and an MTT assay was conducted after 48 h to evaluate the cells’ sensitivity to PSD. (B): Cell proliferation was assessed using an EDU assay. The cells were stained with DAPI (blue) to visualize the nuclei, and proliferating cells were identified by the presence of green fluorescence (EDU). The number of EDU-positive cells per field was quantified from multiple fields, and cell proliferation was calculated relative to the total cell count. Scale bars represent 20 μm. (C): Apoptosis was detected through Hoechst 33,342 and JC-1 staining. The nuclei were stained blue with Hoechst 33,342. Apoptotic changes were indicated by increased fluorescence intensity, nuclear pyknosis, and fragmentation. The JC-1 staining assessed apoptosis by measuring the mitochondrial membrane potential. In viable cells, JC-1 aggregated in the mitochondrial matrix, producing red fluorescence. However, in early apoptotic cells, JC-1 existed as monomers and emitted green fluorescence. The ratio of green-stained cells to the total cell count was quantified from multiple fields. Scale bars: Hoechst 33,342 staining (100 μm), JC-1 staining (20 μm). The experiments were repeated three times