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. 2020 Jan 21;9(2):295. doi: 10.3390/jcm9020295

Figure 4.

Figure 4

Inhibin beta A subunit (INHBA) as the major downstream effector in LIF increases oral cancer progression. (A) Heatmap of mRNA expression profile in Cal27/pLKO and Cal27/shLIF stable clones. (B) Gene set enrichment analysis (GSEA) showed the enrichment of metastatic genes in Cal27/pLKO versus Cal27/shLIF cells. (C) Real-time PCR analysis of BUB1, BIRC5, INHBA, CD44, and UBE2C mRNA expression in Cal27/pLKO and Cal27/shLIF cells (** p < 0.01). (D) Reverse transcription PCR analysis of INHBA mRNA expression in CA9-22 and HSC3 cells transiently transfected LIF-expressed or shLIF plasmids. (E) Cal27/pLKO and Cal27/shLIF stable clones were seeded and transiently transfected with 3 µg of control plasmid or various concentrations of INHBA plasmids and incubated for 48 h, then subcultured in a Boyden chamber overnight. Cell motility toward the lower face of the filter was observed and quantified (** p < 0.01). (F) Western blot analysis of INHBA protein expression in CA9-22 cells after they were treated with rLIF. β-actin was used as an internal control.