Journal: International journal of cancer

Article Title: MED15, encoding a subunit of the mediator complex, is overexpressed at high frequency in castration-resistant prostate cancer.

doi: 10.1002/ijc.28647

Figure Lengend Snippet: Figure 5. MED15 knockdown and proliferation in PCa cells. Correlation of MED15 expression with tumor proliferation markers Ki67 and PHH3 (a). Expression of MED15 in PC3 (b) and DU145 (c) nuclear extracts by Western blot. SiRNA mediated MED15 knockdown in DU145 and PC3 cell lines by Western blot (d) and IHC (e). MTT proliferation assay of PC3 cells (f). *p < 0.05, **p < 0.01, ***p < 0.001. Similarly, proliferation assay of DU145 cells (g). *p < 0.05, **p < 0.01, ***p < 0.001. SiRNA mediated MED15 knockdown in LNCaP cell line (h). Prolif- eration assay of LNCaP cells treated with DHT (i). *p < 0.05, **p < 0.01, ***p < 0.001. ***p < 0.001, Pearson. ShRNA stable knockdown of MED15 in the DU145 cell line (j). Proliferation assay of DU145 cells with stable knockdown of MED15 compared to the scrambled control (k). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar, 10 mm. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: Cell lysates and nuclear extracts of PC3 (sc-2152, Santa Cruz, CA) and DU145 (sc-24960, Santa Cruz, CA) were analyzed by immunobloting using MED15 rabbit polyclonal antibody (Proteintech, Chicago, IL).

Techniques: Knockdown, Expressing, Western Blot, Proliferation Assay, shRNA, Control

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a SLFN11 expression in normal tissues, arrows indicate (from left to right); negative breast lobular cells, positive (red) and negative (blue) prostatic cells, positive lymphocytes, positive bronchial epithelial cells, negative colon epithelial cells. Negative Langerhans islets (blue circle) and positive cluster of acinar cells (red circle) also shown. b SLFN11 prevalence in various cancer types from multi-tumour tissue microarrays (TMAs), expression of SLFN11 shown as percent of positive tumour cells when internal control staining was acceptable. Median given (green) for each cancer type. c Example images of SLFN11 IHC staining in head and neck, thyroid, lymphoma and prostate cancers (Ca). d , e Breakdown of the SLFN11 protein levels by: d breast cancer subtype; oestrogen receptor-positive (ER + ), human epidermal growth factor receptor 2 (HER2 + ), triple-negative breast cancer (TNBC), invasive lobular breast cancer (ILC) and e thyroid cancer subtype; papillary and follicular. f – h SLFN11 expression in colorectal cancer (CRC) patients ( n = 144) subcategorised by clinical patient information: f presence of metastasis or not; g tumour grade, h disease stage. Data shown as median ± interquartile range. * P value <0.05 by Mann–Whitney test. Black arrows are stromal and endothelial cells. Scale bars at 100 µm.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Expressing, Control, Staining, Immunohistochemistry, MANN-WHITNEY

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a – c SLFN11 sub-clonal expression pattern demonstrated by IHC of a a CRC TMA core, b resected serous ovarian cancer tissue, c resected breast cancer tissue. A magnified image of the high- and low SLFN11 subclones are shown for each. Black arrows indicate stromal and endothelial cells used as a positive internal control. d RNA in situ hybridisation (ISH) of SLFN11 showing expression of SLFN11 RNA transcripts in the breast cancer SLFN11 subclones. e SLFN11 sub-clonal expression correlated to Ki67 expression. Line indicates patient-matched subclones. * P value < 0.05 paired t test. Scale bars at 50 µm unless otherwise stated.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Expressing, Control, In Situ, Hybridization

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a Image of a SCLC patient tumour demonstrating sub-clonal SLFN11 expression, areas from the high- and low-expressing subclones are magnified. The black arrow indicates SLFN11-positive stromal cells in the negative sub-clone. Scale bars at 100 µm unless otherwise stated. b SLFN11 prevalence by IHC in SCLC patients, median in green. c Bimodal prevalence of SLFN11 in the SCLC cohort ( n = 124). d SLFN11 expression in SCLC subcategorised by disease stage. Black; SLFN11 monoclonal expression, Red; SLFN11 sub-clonal tumours. Median ± interquartile range shown. e – h Kaplan–Meier analysis of SCLC patients categorised by SLFN11 expression; SLFN11 high (H-score >122) compared to low (H-score ≤122). e Progression-free survival (PFS) and f overall survival (OS) in months for patients ( n = 24) that received first-line platinum (carboplatin or cisplatin) plus etoposide. g PFS and h OS in months for 45 and 57 patients, respectively, that received any chemotherapy. Events table with patients at risk shown for each timepoint.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a Prevalence of SLFN11 by IHC H-score in serous ovarian cancer patient biopsies ( n = 151), cut-off depicting the high and low SLFN11 subgroups shown, b representative images of high and low SLFN11 tumours. Black arrow indicates SLFN11-positive stromal cells used as an internal control. Scale bars at 100 µm. c SLFN11 H-score of patients divided by extreme good and poor clinical responders to carboplatin and paclitaxel doublet therapy. Median ± interquartile range shown. d SLFN11 IHC protein expression by H-score compared to NanoString quantified SLFN11 gene expression in the HGSOC patients. Dotted line indicates gene expression threshold that distinguishes positive from negative SLFN11 patients. Patients with sub-clonal SLFN11 indicated by a red dot. e , f Kaplan–Meier survival curves of ( e ) progression-free interval (PFI) and ( f ) overall survival (OS) of high (>30 H-score SLFN11) vs. low SLFN11 ( ≤ 30 H-score SLFN11) subgroups in the 34 patients with high-grade serous ovarian cancer treated with carboplatin and paclitaxel doublet. Events table with patients at risk shown for each timepoint.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Control, Expressing, Gene Expression

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a Cox proportional hazards model of progression-free survival (PFS) of high (>30 H-score SLFN11) vs. low SLFN11 ( ≤ 30 H-score SLFN11) subgroups in platinum-sensitive serous ovarian carcinoma patients treated with either placebo or olaparib. b Overall survival (OS) of high vs. low SLFN11 in the same study. Events table with patients at risk shown for each timepoint.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques:

Journal: British Journal of Cancer

Article Title: Retrospective analysis of Schlafen11 (SLFN11) to predict the outcomes to therapies affecting the DNA damage response

doi: 10.1038/s41416-021-01560-1

Figure Lengend Snippet: a – d Cox proportional hazards model by SLFN11 high (>30 H-score SLFN11) vs. low SLFN11 ( ≤ 30 H-score SLFN11) subgroups in platinum-sensitive serous ovarian carcinoma patients treated with either placebo or olaparib for ( a ) progression-free survival (PFS) in BRCA wild-type patients, b PFS in BRCA mutant patients, c Overall survival (OS) in BRCA wild-type patients, d OS in BRCA mutant patients. Events table with patients at risk shown for each timepoint. VUS BRCAm patients included in the BRCAwt group.

Article Snippet: DU145 cells (DSMZ) and DU145 CRISPR-Cas-9 SLFN11 knock-out cell line generated by AstraZeneca (Oncology Bioscience UK) cultured in EMEM media with 10% foetal bovine serum (ATCC).

Techniques: Mutagenesis