Fig. 5: Grpr expression in female mice by a Cdh1/Ctnnb1/Esr1/Grpr amplification loop.
From: Targeting GRPR for sex hormone-dependent cancer after loss of E-cadherin
a, Enrichment of mouse ChIP–seq signatures in ∆Ecad female-specific H3K27ac peaks located at gene bodies or promoters. b,c, Quantitative PCR with reverse transcription (RT–qPCR) of Nkd1, Esr1 and Grpr in Ecad mouse 1014 melanoma cells after siScr versus siCdh1 in the presence of β-catenin (bcat; b) and pcDNA3 versus β-catenin transfection (c). d, Heatmap of sex-hormone receptor expression in Ecad and ∆Ecad melanoma cell lines, annotated with Cdh1 and Grpr levels. M, male; F, female e, Western blot of E-cadherin and ERα in mouse melanoma cell lines. Actin was used as a loading control. f,g, Effect of Esr1 knockdown (f) or overexpression (g) on GRPR in mouse and human melanoma cells. h, Grpr expression after Cdh1 and/or Esr1 knockdown in Cdh1+ mouse melanoma cells. i, Western blot analysis of Ecad and ERα after siScr, siCdh1 or siESR1 in 1014 cells. Actin was used as a loading control. j, Western blot of ECAD in 501mel cells with and without GRPR expression and GRP treatment. k, Quantification of lung metastases by stereomicroscopy and RT–qPCR for Cre markers in lungs. The log-normalized expression values were compared by two-sided Student’s t-test (two groups) or by analysis of variance (ANOVA) corrected by Tukeys’s test (multiple groups). Metastasis burden was assessed by Fisher’s exact test adjusted by the Bonferroni method. Variation of the Cre expression was assessed by two-sided Mann–Whitney tests adjusted for multiple comparisons by a Benjamini–Hochberg test. Data are shown as mean ± s.d. At least three independent biological replicates were performed for each experiment. Ful, Fulvestrant.
