Journal: Molecular Medicine Reports

Article Title: Reoxygenation induces reactive oxygen species production and ferroptosis in renal tubular epithelial cells by activating aryl hydrocarbon receptor

doi: 10.3892/mmr.2020.11679

Figure Lengend Snippet: AhR activation status does not affect Nrf2 activation or transcriptional activity. RPTECs were cultured under ctrl conditions or subjected to Reox with or without the AhR inhibitor CH223191. (A) Representative western blots of Nrf2 levels (corresponding to its activation status) and the expression of the Nrf2 transcriptional targets xCT (SLC7A11) and SOD-3. (B-D) Statistical analysis of the western blots. Neither Reox nor CH223191 affects Nrf2 activity, or the expression of xCT and SOD-3. Data are presented as the mean ± SEM of six independent experiments. AhR, arylhydrocarbon receptor; Nrf2, nuclear factor erythroid 2-related factor 2; xCT, cystine-glutamate antiporter; SOD-3, superoxide dismutase; RPTEC, renal proximal tubular epithelial cell; ctrl, control; Reox, reoxygenation.

Article Snippet: Primary antibodies were specific for AhR (1:200; cat. no. sc-133088; Santa Cruz Biotechnology, Inc.), cytochrome P450 family 1 subfamily A member 1 (CYP1A1; 1:500; cat. no. sc-25304; Santa Cruz Biotechnology, Inc.), Nrf2 (1:1,000; cat. no. TA343586; OriGene Technologies, Inc.), superoxide dismutase 3 (SOD-3; 1:100; cat. no. sc-271170; Santa Cruz Biotechnology, Inc.), cystine-glutamate antiporter (xCT, also known as SLC7A11; 1:1,000; cat. no. ANT-111; Alomone Labs), HIF-1α (1:500; cat. no. sc-10790; Santa Cruz Biotechnology, Inc.), LDH-A (1:1,000; cat. no. 2012; Cell Signaling Technology, Inc.), activated cleaved caspase-3 (CC3; 1:1,000; cat. no. ab13847; Abcam) and β-actin (1:2,500; cat. no. 4967; Cell Signaling Technology, Inc.).

Techniques: Activation Assay, Activity Assay, Cell Culture, Western Blot, Expressing

Journal: bioRxiv

Article Title: L-asparaginase treatment induces a tumor metabolic plasticity and reveals a vulnerability to PARP1/2 inhibitor Olaparib, in B-cell lymphomas

doi: 10.1101/2025.10.24.683536

Figure Lengend Snippet: a. Relative ROS levels in Eµ- Myc (#688) cells cultivated for 15 hours in Asn-free medium, supplemented with (+) or without (-) Asn (0.37 mM) and ASNase (0.003 IU/ml). Prior incubation with the CellROX probe, cells were pretreated (+) or not (-) with N-acetyl-L-cysteine (NAC, 10 mM) for 1 hour, followed by analysis by flow cytometry. Data are expressed as the Median Fluorescence Intensity (MFI) normalized to the control condition (+Asn) and presented as mean ± SD (n=4 experiments). P-value from 2-way Anova followed by Tukey’s test. b. GSSG/GSH ratio in Eµ- Myc (#688) cells incubated for 24 hours in Asn-free medium, supplemented (+) or not (-) with Asn (0.37 mM) for 24 hours. Data are expressed as mean ± SD (n= 4 biological replicates). P-value from t-test. c. Total protein extracts prepared from whole Eµ- Myc cells cultivated for 24 hours as described in a. were immunoblotted for NRF2 and ATF4. ERK2, loading control. Immunoblots shown are representative of 2 independent experiments for each Eµ- Myc line. d. Relative quantification of indicated protein levels expressed in Eµ- Myc #506 (square, n=2 independent experiments) and in Eµ- Myc #688 (triangle, n=2 independent experiments) cells. Data are normalized to the control condition +Asn and represented as mean ± SD. P value from t-test. e. Relative mRNA expression levels of Gclc , Gclm and Nqo1 normalized to Rplp0 in Eµ- Myc (#688) cells after 24 hours incubation under the conditions described in a. Data are expressed as mean ± SD (n= 3 experiments). P-value from t-test. f. g. Relative ROS levels in Eµ- Myc (#688) cells treated for 20 hours with (+) or without (-) Asn (0.37 mM) and/or the PHGDH inhibitor BI-4916 (10 µM) in Asn-free medium ( f. ) or with (+) or without ((-), DMSO) ASNase (0.003 IU/ml) and/or BI-4916 (10 µM) in Asn-containing medium ( g. ). Data are represented as MFI of CellROX green probe, normalized to control condition (+Asn) and expressed as mean ± SD (n= 4 experiments). P-value from 2-way Anova followed by Tukey’s test. h. Percentage of dead Eµ- Myc (#506) cells (DAPI+) treated with (+) or without (-) DMSO, ASNase (0.003 IU/ml), BI-4916 (7.5 µM) and/or NAC (5 mM) for 48 hours in Asn-containing medium. Data are expressed as mean ± SD (n= 4 experiments). P-value from 2-way Anova followed by Tukey’s test. i. Heatmap representation of mRNA expression levels for the indicated genes, determined by RNAseq from Eµ- Myc cells isolated from 12 individual transgenic Eµ- Myc Tg/+ mice. Expression values are shown as log 2 (fpkm). j. As in i. from BCL harvested from Eµ- Myc cells-bearing mice treated with Vehicle or ASNase (n=1 tumor/group). Expressions values are shown as log 2 (fpkm). k. slc7a11 mRNA expression levels (transcript per million) in selected tumor entities available on the TCGA database. HNSC, Head and Neck squamous cell carcinoma; LUSC, Lung squamous cell carcinoma; LUAD, Lung adenocarcinoma; LGG, Brain Lower Grade Glioma; GBM, Glioblastoma multiforme; STAD, Stomach adenocarcinoma; SKCM, Skin Cutaneous Melanoma; COAD, colon adenocarcinoma; DLBCL, Diffuse Large B-cell Lymphoma; LAML, Acute Myeloid Leukemias. l. Whole-cell lysates prepared from Eµ- Myc (#506) cells stably expressing either control vector (CTL) or the murine SLC7A11-encoding vector (SLC7A11 OE), were immunoblotted for the indicated proteins. ERK2, loading control. Relative quantification below (normalized to the SLC7A11 OE condition). m. Percentage of dead control (CTL) and SLC7A11 overexpressing Eµ- Myc (#506) cells (DAPI+) treated with (+) or without (-) DMSO, ASNase (0.003 IU/ml) and/or BI-4916 (7.5 µM) for 48 hours in Asn-containing medium. Data are expressed as mean ± SD (n= 5 experiments). P-value from 2-way Anova followed by Tukey’s test. ns , not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Article Snippet: Relative mRNA levels were obtained by real-time quantitative PCR (qPCR) on the StepOneTM Real-Time PCR System (Thermo Fisher Scientific) using the TaqMan assay primer set (Thermo Fisher Scientific) for murine Phgdh (Mm01623589_g1), Psat1 (Mm01613328_g1), Psph (Mm01197775_m1), Gls (Mm01257297_m1), Gclc (Mm00802658_m1), Gclm (Mm01324400_m1), Nqo1 (Mm01253561_m1), slc7a11 (Mm00442530_m1), slc6a9 (Mm00433662_m1), Asns (Mm01137310_g1), and the TaqMan Universal PCR Master Mix (4304437, Fisher Scientific) according to manufacturer’s instructions.

Techniques: Incubation, Flow Cytometry, Fluorescence, Control, Western Blot, Quantitative Proteomics, Expressing, Isolation, Transgenic Assay, Stable Transfection, Plasmid Preparation

Journal: bioRxiv

Article Title: Oxymatrine Induces Ferroptosis in MKN28 Gastric Cancer Cells

doi: 10.1101/2025.11.22.689973

Figure Lengend Snippet: Effect of oxymatrine on the expression of ferroptosis-related proteins in MKN28 cells. Western blot analysis of ACSL4, GPX4, and SLC7A11 protein expressions in each group.

Article Snippet: Oxymatrine (Shanghai Aladdin Biochemical Technology Co., Ltd.); RPMI 1640 cell culture medium (Gibco); penicillin-streptomycin double antibody (Gibco); CCK-8 kit (Shanghai Yeasen Biotechnology Co., Ltd.); iron ion detection kit (Nanjing Jiancheng Bioengineering Institute); protein extraction kit (Beyotime Biotechnology Co., Ltd.); GPX4, SLC7A11, and Nrf2 antibodies were purchased from Proteintech Group, Inc.

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: A) Quantitative real-time PCR analysis of SLC7A11 and SLC3A2 expression in total RNA extracts from normal pancreatic fibroblasts (isolated from n=8 patients with benign pancreatic conditions) and CAFs (isolated from n=10 PDAC patients). Bars show mean+s.e.m., circles indicate independent replicates (*p≤0.05, student t-test). B) Western blot of SLC7A11 in total protein extracts from PDAC cells and human CAFs (Cell lines 1-6). α-tubulin was used as a loading control. C) Immunoflourescence for DAPI, αSMA (CAF marker) and SLC7A11 in a human PDAC tissue specimen obtained through the Australian Pancreatic Cancer Genome Initiative (APGI). (D-G) Human PDA tissue microarrays obtained through the APGI (International Cancer Genome Consortium cohort) were stained for SLC7A11 by immunohistochemistry. D) Samples selected as references for scoring (0,1,2,3) for tumour and stromal compartments are shown (insets show magnified view of cells). Scores of 0-1 were classified as low SLC7A11 expression (“Tumour low ” and “Stroma low ”), scores of 2-3 were classified as high SLC7A11 expression (“Tumour high ” and “Stroma high ”). E-G) Kaplan-Meier survival curves showing the correlation between SLC7A11 expression in tumour cells (E), stroma (F), or a combination of both (G) with overall patient survival (days survived post-diagnosis). Patients that were deceased due to other causes or that were still alive were censored (shown as black ticks on each line graph). Total patient numbers for each group are indicated in the graph keys. Asterisks indicate significance based on (F) multivariate analysis (G) univariate Log-Rank test (*p≤0.05). H) Representative photos of Tumour low Stroma low , Tumour high Stroma low , Tumour low Stroma high , and Tumour high Stroma high groups. Scale bars in all photos = 100μm.

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Isolation, Western Blot, Marker, Staining, Immunohistochemistry

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: A) Cell proliferation (based on cell counting kit 8 absorbance) of CAFs 72h post-transfection with non-silencing siRNA (ns-siRNA) or SLC7A11-siRNA pool (pool of 4 siRNA sequences). Asterisks indicate significance (****p≤0.0001; n=6, student t-test). B) Cell proliferation (cell counting kit 8 absorbance) of CAFs treated with sulfasalazine (SSZ) ± 66μM 2-mercaptoethanol (2-ME), as a % of controls. Circles indicate replicates, asterisks indicate significance (***p≤0.001, ****p≤0.0001; One-way ANOVA). C) Live cell counts of CAFs (as a fraction of controls) treated with erastin for 48h. Asterisks indicate significance (****p≤0.0001, n=4; student t-test). D-E) Radiolabelled cystine uptake as a fraction of ns-siRNA (72h post-transfection) or untreated control cells (48h post-treatment with SSZ). Asterisks indicate significance (*p≤0.05; student t-test). F-G) Intracellular glutathione levels as assessed by colorimetric assay, and as a fraction of ns-siRNA (72h post-transfection) or untreated control cells [16h post-treatment with SSZ±N-acetyl-cysteine (NAC)]. Asterisks indicate significance (*p≤0.05, **p≤0.01, n=6; One-way ANOVA). H-I) Intracellular oxidative stress in the presence or absence of tert-butyl hydroperoxide (tBHP; oxidative stress), as measured by CellROX staining and flow cytometry (as a fraction of ns-siRNA + 0 μM tBHP). Asterisks indicate significance (ns=not significant, **p≤0.01, n=3; One-way ANOVA). Circles in all graphs indicate replicates, lines and bars in all graphs represent mean±s.e.m.

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Cell Counting, Transfection, Colorimetric Assay, Staining, Flow Cytometry

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: A) Live cell counts of CAFs 72h post-transfection with non-silencing-siRNA (ns-siRNA), SLC7A11-siRNA pool or SLC7A11-siRNA single sequence (SLC7A11-siRNA single seq) and 24h post-treatment with tert-butyl hydroperoxide (tBHP). Circles indicate replicates, asterisks and hashes indicate significance (*p≤0.05, ** p≤0.01, **** p≤0.0001; # p≤0.05, ## p≤0.01, relative to ns-siRNA of the same tBHP concentration; n=4; One-way ANOVA). B) Frequency of AnnexinV+DAPI positive (apoptotic) cells, as a fraction of ns-siRNA+0μM tBHP controls, 72h post-transfection and 24h post-tBHP treatment. Circles indicate replicates, asterisks indicate significance (ns=not significant, **p≤0.01, ***p≤0.001; One-way ANOVA). C) Glutathione peroxidase activity of CAFs treated with erastin (9h) as a % of controls. Circles indicate replicates, asterisks indicate significance (*p≤0.05; n=4; student t-test). D) Live cell counts of CAFs 24h post-treatment with 40 μM erastin ± 2μM ferrostatin. Circles indicate replicates, asterisks indicate significance (*p≤0.05, n=4; One-way ANOVA). E) Live cell counts (trypan blue exclusion) of CAFs stably expressing scramble-shRNA or SLC7A11-shRNA seq 1, 72h post-seeding and 24h post-treatment with tBHP. Circles indicate replicate experiments. Asterisks represent significance (**p≤0.01, ***p≤0.001, ****p≤0.0001; n=3; One-way ANOVA). F) As per C, except CAFs stably expressed scramble-shRNA or SLC7A11-shRNA sequence 1 and were treated with 40uM tBHP for 9h, instead of erastin (**p≤0.01; n=3; One-way ANOVA). G-H) β-galactosidase positive cells (senescent cells) as a fraction of total cells (mean+s.e.m.): (G) 72h after transfection with control siRNA (ns-siRNA) or SLC7A11-siRNA or (H) 48h post-treatment with SSZ. Circles indicate replicates, asterisks indicate significance (**p≤0.01; G: n=4, student t-test; H: n=3, One-way ANOVA). Bars and lines in all graphs are mean±s.e.m. Replicate numbers in all panels refer to experiments performed using independent CAF cells isolated from different PDAC patients.

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Transfection, Sequencing, Concentration Assay, Activity Assay, Stable Transfection, Expressing, shRNA, Isolation

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: A-B) Schematic diagram of 3D co-culture spheroid outgrowth assay and quantification of 3D co-culture spheroid outgrowth post-transfection with control-siRNA (ns-siRNA) or SLC7A11-siRNA pool. Labels: PDAC ns CAF ns = non-silencing controls; PDAC ns CAF slc = SLC7A11 knockdown in CAFs only; PDAC slc CAF ns = SLC7A11 knockdown in PDAC cells only; PDAC slc CAF slc = SLC7A11 knockdown in both PDAC cells and CAFs. Representative photos are shown above each bar with the core circled in white dashed lines and outgrowth in red dashed lines (bars in photos = 300μm). Circles indicate replicates, lines indicate mean±s.e.m., asterisks indicate significance (ns=not significant, *p≤0.05, **p≤0.01, ***p≤0.001; One-way ANOVA). C) Schematic diagram of 3D co-culture growth assay using stable shRNA cell lines and D) representative photos (bars in photos = 200μm) and quantification of 3D co-culture spheroid growth. Labels: PDAC scr CAF scr = scramble-shRNA controls; PDAC scr CAF slc = SLC7A11-shRNA seq 1 in CAFs only; PDAC slc CAF scr = SLC7A11-shRNA seq 1 in PDAC cells only; PDAC slc CAF slc = SLC7A11-shRNA seq 1 in both PDAC cells and CAFs. Circles indicate replicates, lines indicate mean±s.e.m., asterisks indicate significance (ns=not significant, *p≤0.05, **p≤0.01, ****p≤0.0001, n=4-6; One-way ANOVA). Replicate numbers in panel B refer to independent experiments performed using MiaPaCa-2 combined with CAF cells isolated from different PDAC patients. Replicate numbers in panels D refer to replicate spheroids performed using MiaPaCa-2 PDAC cells combined with an immortalised CAF line.

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Co-Culture Assay, Transfection, Growth Assay, shRNA, Isolation

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: A) Schematic diagram of assay and representative photos of collagen plugs contracted by CAFs transfected with control-siRNA (ns-siRNA) or SLC7A11-siRNA pool over 6 days are shown. The line graph shows the average area of contracted plugs at the indicated time points (mean±s.e.m.; *p≤0.05, n=4; One-way ANOVA). B-E) Analysis of collagen content in collagen plugs contracted by CAFs transfected with ns-siRNA or SLC7A11-siRNA at assay endpoint. B) Average picrosirius red signal. Circles indicate replicates, lines indicate mean±s.e.m., asterisks indicate significance (**p≤0.01, n=4; student t-test). Representative images of picrosirius red staining are shown. C) Average total birefringence. Circles indicate replicates, lines indicate mean±s.e.m., asterisks indicate significance (*p≤0.05, n=4; student t-test). Representative birefringence images are shown. D) Average % of total birefringence that was high (red-orange), medium (yellow) and low (green). Circles indicate replicates, lines indicate mean±s.e.m., asterisks indicate significance (*p≤0.05, n=4; student t-test). E) Left graph shows the average maximum second harmonics generation (SHG) signal detected by two-photon confocal microscopy of collagen plugs. Representative SHG images are shown. Right graph shows the average correlation based on GLCM analysis of SHG maximum intensity projections. Circles indicate biological replicates, lines indicate mean±s.e.m., asterisks indicate significance (ns=not significant, *p≤0.05, n=4; student t-test). Replicate numbers in all panels refer to independent experiments performed using independent CAF cells isolated from different PDAC patients.

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Transfection, Staining, Confocal Microscopy, Isolation

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: A) Quantification of Pancreatic Intraepithelial Neoplasia (PanINs) 1A-3 from KC mice (n=7) and KC mice with SLC7A11 conditional KO under Pdx1 -promoter (KC Slc7a11 fl/fl ; n=7) at 70 days of age (mean±s.e.m.). B) Kaplan-Meier analysis showing survival percentage of KPC (n=26) and KPC Slc7a11 fl/fl mice (n=24) mice. C) Representative photos of KPC and KPC Slc7a11 fl/fl tumour sections probed for αSMA (brown). The quantification of αSMA staining is shown in the graph (mean±s.e.m.), based on ImageJ analysis of representative regions from each tumour section (n=5 mice per group). Scale bars = 400μm. D) Representative photos of KPC and KPC Slc7a11 fl/fl tumour sections. The quantification of picrosirius red staining is shown in the bar graph (mean±s.e.m.), based on ImageJ analysis of representative regions from each tumour section. Scale bars = 400μm. Asterisks indicate significance (*p≤0.05; n=5 mice per group; student t-test). E-F) Live cell counts (mean±s.e.m.) of (E) KPC PDAC cells and (F) KPC CAFs 72h post-transfection with control-siRNA (ns-siRNA) or mouse SLC7A11-siRNA pool (SLC7A11 pool). Asterisks indicate significance (*p≤0.05, **p≤0.01; n=3; one-way ANOVA).

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Staining, Transfection

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: All orthotopic tumours were co-injections of PDAC cells and CAFs. A) Orthotopic pancreatic tumours were treated with STAR nanoparticles + control-siRNA or SLC7A11 siRNA single sequence (SLC7A11 single seq) in the regimen shown. Representative photos of immunohistochemistry for SLC7A11 in tumour tissue at the model endpoint are shown. Graph shows optical density (staining intensity) calculated from average pixel intensity measurements from 3 representative images per tumour, using ImageJ. Circles indicate individual mice, lines indicate mean±s.e.m., asterisks indicate significance (*p≤0.05; One-way ANOVA). B) Treatment regimen for therapeutic model analysed in panels (C-D). Circles and triangles in all dot plots in panels C-D represent individual mice. C) Tumour volume at therapeutic model endpoint, as assessed by calliper measurement ex vivo (mean±s.e.m.). Asterisks indicate significance (*p≤0.05; One-way ANOVA). D) Representative photos of metastases confirmed by H&E staining following detection at model endpoint by ex vivo luminescence imaging of organs. Graph shows metastatic sites per mouse (mean±s.e.m.) for each treatment group. Scale bars in all figures = 200μm.

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Sequencing, Immunohistochemistry, Staining, Ex Vivo, Imaging

Journal: bioRxiv

Article Title: Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma determine response to SLC7A11 inhibition

doi: 10.1101/2020.07.12.199638

Figure Lengend Snippet: A) Representative photos of tumour sections probed for αSMA (brown). The quantification of αSMA staining is shown in the graph (mean+s.e.m.), based on ImageJ analysis of representative regions from each tumour section. Asterisks indicate significance (*p≤0.05; Control-siRNA, n=7; SLC7A11-siRNA single seq, n=5; student t-test). B) Representative photos of picrosirius red and methyl green stained tumour sections. The quantification of picrosirius red staining is shown in the bar graph (mean+s.e.m.), based on ImageJ analysis of representative regions from each tumour section. Asterisks indicate significance (*p≤0.05; Control-siRNA, n=8; SLC7A11-siRNA single seq, n=5; student t-test). C) Polarised light analysis of representative regions from picrosirius red stained specimens. Representative photos are shown. Left bar graph shows total birefringence (mean+s.e.m.; Control-siRNA, n=8; SLC7A11-siRNA single seq, n=5). Right bar graph shows the average frequency (mean+s.e.m.; Control-siRNA, n=8; SLC7A11-siRNA single seq, n=5) of low, medium and high birefringence collagen fibrils (higher birefringence = denser fibril). ns = not significant (student t-test). D) Representative photos of CD31-stained tumour sections. Red arrows indicate open blood vessels. The bar graph shows the fraction of CD31-positive blood vessels that were open (mean+s.e.m.), based on ImageJ analysis of representative regions from each tumour section. Asterisks indicate significance (*p≤0.05; Control-siRNA, n=8; SLC7A11-siRNA single seq, n=5; student t-test). Fields of view used for analyses in all panels, provided an average area coverage of 13% of the total tumour section (excluding necrotic regions). All circles in graphs represent individual mice. All scale bars in photos = 100μm.

Article Snippet: MiaPaCa-2 cells and hTERT-immortalised CAFs were then transduced with lentiviral constructs expressing scramble shRNA, SLC7A11-shRNA sequence 1 (Origene, Cat. TL309282).

Techniques: Staining