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. 2022 Jun 30;12:939969. doi: 10.3389/fonc.2022.939969

Figure 6.

Figure 6

LIFR antagonist EC359 hinders cell cycle progression, increases apoptosis rate in MKN45 cells and inhibits EMT process. (A) Dose–response curve of EC359 (25, 50, 100, and 1,000 nM) determined using MTS assay on MKN45 cells (n = 10). MKN45 cells were serum-starved and triggered with LIF (10 ng/ml), EC359 (100 nM), and LIF + EC359 for 48 h. Cell cycle phase analysis was performed by Ki-67/DAPI staining through IC-FACS. Data shown are follows: percentage of (B) from left to right cell in G0-G1 cell cycle phases, S-G2-M cell cycle phases, and ratio between % G0-G1 and % S-G2-M. (C) Percentage of apoptotic cells. (D) Representative IC-FACS showed cell cycle fraction and apoptosis rate in NT, LIF (10 ng/ml), EC359 (25 nM), and LIF + EC359. Results are the mean ± SEM of three samples for group (* represents statistical significance versus NT, and # versus LIF, p < 0.05). Relative mRNA expression of (E) the proliferation marker C-Myc and EMT markers (F) E-Cadherin, (G) Snal-1, and (H) vimentin. Each value is normalized to Gapdh and is expressed relative to those of positive controls, which are arbitrarily settled to 1. Results are the mean ± SEM of five samples per group (* represents statistical significance versus NT, and # versus LIF, p < 0.05).