Fig. 5. BACH1ʼs nonmonotone invasion landscape is independent of native BACH1.
a, Western blot examination of native BACH1 protein level dose response in low-noise mNF-BACH1 and mNF-GFP MB231 samples. Native BACH1 bands are highlighted in the red rectangle. b, Quantitation of native BACH1 protein level dose response in low-noise mNF-BACH1 and mNF-GFP MB231 samples using grayscale normalization to internal reference β-tubulin in a. c, mRNA level dose responses of truncated BACH1 isoform, BACH1t, in low-noise mNF-BACH1 MB231 clones. BACH1t is transcribed only from the native copy of BACH1 gene. One-way ANOVA with Tukey’s multiple comparisons tests between each dose and uninduced controls, n = 3, *P < 0.05, **P < 0.01. d, Invasiveness significantly decreased due to BACH1 knockout (KO) compared to the wild-type parental population. Invasiveness can be rescued by transient overexpression from the induced mNF-BACH1 circuit but not by BACH1t overexpression. Two-tailed t-test between BACH1-KO sample and every other condition, n = 3, *P < 0.05, **P < 0.01. e,f, Dose responses of MFI (e) and CV (f) for a selected MB231 BK monoclone with AAVS1 site-specifically integrated mNF-BACH1 circuit relative to the low-noise mNF-BACH1 (BL) clones (n = 3). g, Invasiveness of a selected MB231 BK + mNF-BACH1 circuit monoclone relative to low-noise mNF-BACH1 (BL) clones over increasing dox concentrations (n = 3). NS, not significant.