Journal: Molecular cell

Article Title: Removal of RTF2 from stalled replisomes promotes maintenance of genome integrity

doi: 10.1016/j.molcel.2017.11.035

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Dried slides were stained with a 1:30000 dilution of SYBR-GOLD and visualized and analyzed by Metasystems CometScan software.

Techniques: Recombinant, Single Cell Gel Electrophoresis, Protein Quantitation, Software

Journal: bioRxiv

Article Title: Progressive plasticity during colorectal cancer metastasis

doi: 10.1101/2023.08.18.553925

Figure Lengend Snippet: a, Gene trends along aligned canonical-non-canonical axes, as in , of the 6 top-ranked fetal-associated human TFs in four patients. TFs are ranked according to their mean correlations of imputed gene expression with the fetal signature score in these patients. b , Lunaphore COMET immunofluorescence imaging of CDX2 (intestinal lineage), PROX1 (fetal state) and CK5 (squamous lineage) (top) and hematoxylin and eosin imaging (bottom) of the KG146P invasion front, showing the relationship between PROX1 expression and the transition from well-differentiated glandular morphology to poorly differentiated state. Arrow indicates a blood capillary completely encased by mucosa invasive cancer cells (scale bar = 500 μm). c , COMET immunofluorescence imaging of two KG146Li liver metastasis fields of view, showing spatially ordered transitions in expression of CDX2 (intestinal lineage), PROX1 (fetal state) and CK5 (squamous lineage) among metastatic tumor cells (scale bar = 1000 μm (top), 100 μm (bottom)). d,e, Organoid initiation capacity (number of organoids formed per 2000 single cells/40 μL matrigel) (d) and organoid outgrowth capacity (average organoid diameter at day 7) (e) of OKG146P and OKG146Li organoids expressing doxycycline- inducible shRNAs targeting Prox1 or control shRNA at 7 days of culture in HISC media supplemented with 2 μg/ml doxycycline for 7 days. (scale bar = 1000 μm; n = 8; one-sided t-test, Benjamini-Hochberg correction, *, p < 0.05, ****, p < 0.0001). f-i, Relative mRNA expression of differentiated intestine and squamous differentiation markers in OKG146P (f) , OKG146Li (g) OK136P (h) and OK136Li (i) organoids expressing doxycycline-inducible shRNAs targeting PROX1 , cultured in HISC media containing 2 μg/ml doxycycline for 7 days, in comparison with shControl organoids. RT-qPCR normalized to GAPDH mRNA expression (n = 4, t-test, Benjamini-Hochberg correction, * indicates p < 0.05). j , Model of cell-state transitions during metastatic tumor progression in CRC. Cancer cells in the primary tumor first enter into an ISC-like state. Cells at the primary tumor invasion front undergo developmental reversion into a highly plastic fetal progenitor-like state, enabling non-canonical differentiation into divergent states, including those resembling neuroendocrine and squamous cells, during metastatic outgrowth. Epithelial injury during tumor dissemination, metastasis and after therapy induces entry into the highly plastic fetal progenitor state, enabling tumor regenerative cells to express gene programs of non-canonical lineages, and dynamically adapt to diverse stresses during metastasis and therapy. Induction of the transcriptional repressor PROX1 serves to restrict non-canonical gene expression in injured normal epithelia, enabling tissue regeneration. PROX1-responsive intestinal lineage restriction is progressively lost during cancer progression, licensing non-canonical differentiation.

Article Snippet: The OME.tif image was exported after background subtraction with Lunaphore COMET Viewer software which was visualized and assessed per channel using Indica Lab Halo software.

Techniques: Expressing, Immunofluorescence, Imaging, shRNA, Cell Culture, Comparison, Quantitative RT-PCR