Journal: PLoS ONE

Article Title: CoREST1 Promotes Tumor Formation and Tumor Stroma Interactions in a Mouse Model of Breast Cancer

doi: 10.1371/journal.pone.0121281

Figure Lengend Snippet: (A) Conditioned media from shcontrol (shCtrl) and shCoREST1 (shCoR #1) MDA-MB-231 cells was incubated with a human angiogenesis antibody array as described in Materials and Methods. Immunoblot images from this screen, performed one time, are shown. (B) Quantification of the relative pixel density on the array for the indicated pro-angiogenic, pro-inflammatory and anti-angiogenic factors secreted by shCtrl and shCoR cell lines (n = 1 experiment). (C) VEGF-A mRNA expression was measured in shCoR cells compared to shCtrl cells in MDA-MB-231 and SUM159 cell lines. (D) CCL2 mRNA expression was measured in the indicated cell lines. Expression levels were detected by RT-qPCR and represented as fold change compared to shCtrl cells. Differences were determined by Student’s t-test (mean±s.d.; n = 3 experiments). (E) Luciferase activity from shCtrl or shCoR #1 MDA-MB-231 cells transfected with VEGF-luc or pMCP-luc and pRL-CMV-Renilla. Luciferase expression was normalized to Renilla, then expressed as fold change compared to shCtrl cells. Differences were determined by Student’s t-test (mean±s.d.; n = 3 experiments).

Article Snippet: 1 ml of conditioned media was used to probe a human angiogenesis antibody array (R&D Systems cat. no. ARY007), according to the manufacturer’s instructions.

Techniques: Incubation, Ab Array, Western Blot, Expressing, Quantitative RT-PCR, Luciferase, Activity Assay, Transfection

Journal: Nature Communications

Article Title: RNA splicing is a key mediator of tumour cell plasticity and a therapeutic vulnerability in colorectal cancer

doi: 10.1038/s41467-022-30489-z

Figure Lengend Snippet: a Representative histological images of mouse colons and tumours stained with hematoxylin and eosin (H&E). Scale bars are 1 mm (2.5x), 500 µm (5x) and 250 µm (10x). Tumours were isolated when mice showed clinical signs of intestinal tumourigenesis. b Quantification of the presence or absence of invasive tumours in mice from the indicated genotypes, with statistical difference calculated using a two-sided Chi-square test, n = 23 vs 34 biologically independent mice. c Box and whisker plot showing the number of invasive tumours as a percentage of the total number of tumours for each mouse for each genotype. The box extends from 25th to 75th centiles, the centre line is median and whiskers extend to minima and maxima, n = 23 vs 34 biologically independent mice. d Representative histological images of p53 fl/fl and p53; fl/fl Srsf1 fl/+ mouse intestines and tumours stained for PROX1 (stem cell marker) and SLC13A2 (differentiation marker). Scale bars are 2.5 mm (1.25x), 500 µm (5x) and 50 µm (40x). e Histoscore and staining strength quantification for PROX1, n = 5 vs 6 biologically independent mice and f SLC13A2, n = 7 vs 5 biologically independent mice. g Schematic depiction of the experimental strategy used to investigate the invasive potential of tumour-derived intestinal organoids. h Representative images of calcein-stained organoids once they had invaded through Matrigel and through the porous membrane at the bottom of the assay well. Scale bar 500 µm. i Quantification of the invasive potential of p53 fl/fl tumour-derived organoids following control or Srsf1 shRNA manipulation, n = 9 vs 9 independent experiments. Data in bar charts are represented as mean and error bars are SD with data analysed with two-tailed, unpaired t -tests, p values are indicated in figure panels. All biological replicates are shown as individual value plots and n > 3. See also Fig. .

Article Snippet: The following antibodies were used: BrdU (BD Biosciences, 347580, pH6, 1/500), PROX1 (R&D systems, AF2727, pH6, 1/100 for human tissue array, 1/200 for mouse sections), SLC13A2 (Atlas antibodies, HPA014963, pH8, 1/100), SRSF1 (Invitrogen/Thermo Fisher Scientific, 32-4600, pH8, 1/10,000), Ki67 (Abcam, ab15580, pH6, 1/2000), Active Caspase-3 (R&D systems, AF835, pH6, 1/800).

Techniques: Staining, Isolation, Whisker Assay, Marker, Derivative Assay, Membrane, Control, shRNA, Two Tailed Test