Journal: BMC Biology

Article Title: A ZEB1-Neon knock-in uncovers traceable dynamics of epithelial-mesenchymal transition in tumors in vivo

doi: 10.1186/s12915-026-02629-0

Figure Lengend Snippet: Endogenous tagging of ZEB1 by mNeonGreen (Neon) with CRISPaint faithfully reports on ZEB1 expression in MDA-MB-231 and MCF10A cells. A Schematic representation of the CRISPaint approach, using a modified donor plasmid for seamless fusion at ZEB1 exon 9, integrating Neon-T2A-puro. sgRNA-directed expected Cas9 cuts in the ZEB1 locus and plasmid are indicated by red arrowheads. B Western blot analysis of successfully targeted MDA-MB-231 after clonal expansion of ZEB1-Neon clones E2, E9, E10, F11, A5, and B6 together with the parental (par) and one control clone (ctrl). Anti-ZEB1 immunoblotting identifies both ZEB1 wildtype and ZEB1-Neon proteins as indicated. C Immunofluorescence staining (IF) using anti-mNeonGreen and anti-ZEB1 antibodies of selected clones from B . Nuclei are stained with DAPI; arrowheads, ZEB1hi cells; open arrow, ZEB1lo cells. D Representative FACS plots and mean fluorescence intensity (MFI) of n = 4 experiments for MDA-MB-231 ZEB1-Neon, par and ctrl clones as in B . E Detection of intrinsic ZEB1-Neon fluorescence and DIC (BF) imaging using spinning-disc confocal imaging of clones from B . F , G Western blot of MCF10A par, two ctrl (E9, E11), and two successfully CRISPaint-targeted clones (B6, D9) in standard culture ( F ) and upon TGFβ treatment for 14 days of par, F8 ctrl, and D9 Zeb1-Neon clones ( G ). For anti-Neon detection in G , samples were run on opposing ends of the same membrane and merged (vertical separation). H IF imaging of F8 ctrl and D9 MCF10A clones with and without TGFβ treatment for 14 days as in C ; arrowheads, ZEB1hi cells; open arrow, ZEB1lo cells. Scale bars, 50 µm ( C , H ) and 20 µm ( E )

Article Snippet: MDA-MB-231 and MCF10A were purchased from American Type Culture Collection (ATCC) and cultured in DMEM (Gibco, 31966021)/10% FBS (Gibco, 10500064) and DMEM/F-12 (Gibco, 31331028)/5% horse serum (Gibco, 16050122)/20 ng/ml EGF (Peprotech, 100–15)/0.5 mg/ml hydrocortisone (Sigma, H0888)/0.1 mg/ml cholera toxin (Sigma, C8052)/10 mg/ml insulin (I9278), respectively, at 37 °C/5% CO 2 in a humidified incubator as described previously [ ].

Techniques: Expressing, Modification, Plasmid Preparation, Western Blot, Clone Assay, Control, Immunofluorescence, Staining, Fluorescence, Imaging, Membrane

Journal: Frontiers in Genetics

Article Title: Identification and validation of a seven-gene metastasis-associated prognostic model in breast cancer

doi: 10.3389/fgene.2026.1770418

Figure Lengend Snippet: Prognostic gene expression in cell lines. (A) RTN1-A, RTN1-B and RTN1-C expression in tumor (n = 1,080) and para-tumor (n = 98) samples from TCGA-BRCA. (B) Ratios of RTN1-A/RTN1, RTN1-B/RTN1 and RTN1-C/RTN1 in tumor and paratumor samples from TCGA-BRCA. (C) mRNA levels of prognostic genes in MCF-10A, MCF-7, SKBR3, MDA-MB-231 and MDA-MB-468 cells (n = 4). (D) Electrophoretogram of the PCR products of CXCL14 (171 bp), RTN1-A (134 bp), and RTN1-C (78 bp). The data are presented as the means ± SDs. P values were calculated via Student’s unpaired t-test. *P < 0.05; **P < 0.01; ***P < 0.01; ****P < 0.0001.

Article Snippet: The human mammary epithelial cell line MCF-10A (RRID: CVCL_0598) and breast cancer lines MCF-7 (RRID: CVCL_0031), SKBR3 (RRID: CVCL_0033), MDA-MB-231 (RRID: CVCL_0062) and MDA-MB-468 (RRID: CVCL_0419) were purchased from Procell.

Techniques: Gene Expression, Expressing

Journal: Frontiers in Genetics

Article Title: Identification and validation of a seven-gene metastasis-associated prognostic model in breast cancer

doi: 10.3389/fgene.2026.1770418

Figure Lengend Snippet: Prognostic gene expression in cell lines. (A) RTN1-A, RTN1-B and RTN1-C expression in tumor (n = 1,080) and para-tumor (n = 98) samples from TCGA-BRCA. (B) Ratios of RTN1-A/RTN1, RTN1-B/RTN1 and RTN1-C/RTN1 in tumor and paratumor samples from TCGA-BRCA. (C) mRNA levels of prognostic genes in MCF-10A, MCF-7, SKBR3, MDA-MB-231 and MDA-MB-468 cells (n = 4). (D) Electrophoretogram of the PCR products of CXCL14 (171 bp), RTN1-A (134 bp), and RTN1-C (78 bp). The data are presented as the means ± SDs. P values were calculated via Student’s unpaired t-test. *P < 0.05; **P < 0.01; ***P < 0.01; ****P < 0.0001.

Article Snippet: MCF-10A cells were maintained in MCF-10A cell complete medium (Procell, CM-0525) (DMEM/F12 + 5% house serum + 20 ng/mL EGF + 0.5 μg/mL hydrocortisone +10 μg/mL insulin + 1% NEAA + 1% P/S).

Techniques: Gene Expression, Expressing