Fig. 4

Protodioscin decreased the ROS level and upregulated HO-1 and NQO1 during myofibroblast transition. Primary mouse lung fibroblasts were co-incubated for 24 h with 10 μM, 30 μM, and 100 μM protodioscin with or without TGF-β1 (10 ng/mL). ROS fluorescence probes were used to visualise the level of ROS by microscopy after incubation (A). The proportion of ROS-positive cells was detected by flow cytometry (B). WB analysis of Nrf2 levels in nuclear proteins in each group (C). WB analyzed the protein levels of HO-1 and NQO1 in each group (D). Determining the mRNA expression levels of HO-1 and NQO1 in each group was undertaken through qRT-PCR (E, F). Expression of data was undertaken in form of mean ± standard deviation, and the experiments were replicated independently on a minimum of three occasions, * P < 0.05; *It is suggestive of statistical significance at the P threshold below 0.05; **Denotes heightened significance at the P threshold below 0.01; ***Reflects substantial significance at the P threshold below 0.001; ****Demonstrates remarkable significance at the P threshold below 0.0001