Figure 2.

EMT-associated phenotypes induced by chronic IL-1β exposure are also memorized. (A) Growth curve of A549 cells with various IL-1β treatments, normalized to the value on day 0 on each condition. (B) A dot plot showing numbers of colonies at day 14 after A549 cells were plated for anchorage-independent growth (AIG). Each dot represents a duplicate well. (C) Apoptotic markers were evaluated 24 hours after A549 cells were treated with chemotherapy agents. Quantification of cleaved PARP (c’-PARP) and Caspase 3 (c’-Caspase 3) was normalized to samples without IL-1β exposure. (D) Surface PD-L1 expression was assessed by flow cytometry. Quantification from 3 replicates is represented in the bar graph. (E) Pathway analysis of EMT-associated biological processes at different time points. (F) A pie chart summarizing EMT memory phenotypes. “A”, acute IL-1β exposure (72 hours); “C”, chronic IL-1β exposure (≥21 days); “M”, EMT memory (≥7 days after IL-1β withdrawal from chronic exposure). Cis, cisplatin; Etop, etoposide; Doxr, doxorubicin. “n.d.”, non-detectable. All results are reported as mean ± SEM. ****P < 0.0001. See also Fig. S2, Tables S1 and S2.