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. 2021 Feb 24;16(2):e0247652. doi: 10.1371/journal.pone.0247652

Fig 6. Harmine treatment does not affect the level of Twist1 transcripts in breast cancer cells.

Fig 6

A, Analysis of the expression of Twist1 and its target genes in BT549 cells. BT549 Twist+ cells were either untreated (BT549+), treated with DMSO (DMSO) or harmine (HR 20μM) for 48 hr, and the levels of Twist1 mRNA were determined by RT-qPCR (see Materials and Methods). BT549 Twist- cells (BT549-) were included as a negative control. The mRNA levels of Twist1 and its target genes Bmi1 (BMI1), Snail (SNAIL) and ZEB2 (ZEB2) were normalized against that of β-actin, tubulin α 1a and GAPDH. B, The expressions of EMT markers N-Cadherin (N-CADHERIN), E-Cadherin (E-CADHERIN) and Vimentin (VIMENTIN) were analyzed with the same samples as in A. C, Analysis of the expression of Twist1 and its target genes in 4T1 cells. The 4T1 Twist+ cells were either untreated (4T1+), transfected with scrambled control siRNA (SIQ), transfected with Twist1 siRNA (SITWIST) (negative control), treated with 60 μM harmine (HR 60 μM) or treated with 120 μM harmine (HR 120 μM). mRNA levels for Twist1 and its target genes Bmi1 (BMI1), Snail (SNAIL) and ZEB2 (ZEB2) were measured by RT-qPCR. All results were normalized against the mRNA levels of β-actin, tubulin α 1a and GAPDH. D, The expressions of EMT markers N-Cadherin (N-CADHERIN), E-Cadherin (E-CADHERIN) and Vimentin (VIMENTIN) were analyzed with the same samples as in C. All data were pooled from three independent experiments. Definitions of statistical significance are * p < .05, ** p < .01, *** p < .001, and **** p < .0001.