Journal: Cancer letters

Article Title: Protective role of epithelial deoxyhypusine synthase in colorectal carcinogenesis caused by deletion of the adenomatous polyposis coli gene

doi: 10.1016/j.canlet.2025.218002

Figure Lengend Snippet: The expression of the genes encoding for the main proteins involved in polyamine metabolism ( A ) was determined using RNA-Seq data interrogated from published studies using the GEO data repository ( B ); we used data from CRC tissues compared to normal tissues in the same patients (#) or to colon tissues from normal individuals; the size of the squares indicates the number ( n ) of patients with CRC in each study. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by the Benjamini & Hochberg test adjusted for false discovery rate. ( C - D ) Tissues sections of a TMA were immunostained for DHPS and representative images of tissues from patients with grade 1 ( n = 7), grade 2 ( n = 33), or grade 3 ( n = 8) CRC, and paired non-tumor tissues (NT, n = 48) are shown in ( C ); scale bar, 50 μm; quantification (0–300) of DHPS + CECs is shown in ( D ); T, tumors. * P < 0.05 and **** P < 0.0001 by mixed-effects analysis and Šídák’s multiple comparisons test. ( E ) Level of DHPS mRNA in organoids generated from NT and T tissues from 5 patients with CRC. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by mixed-effect analysis and Tukey’s multiple comparisons test. ( F ) Kaplan-Meier curve was used to evaluate the prognostic risk of DHPS gene expression. P was determined by Kaplan-Meier log rank test; there was a total of 946 patients, including 568 and 378 with low and high DHPS expression, respectively.

Article Snippet: Slides were then sequentially incubated with i ) the rabbit polyclonal anti-DHPS Ab (Proteintech; 1:1000) overnight at 4°C and EnVision+/HRP Labelled Polymer Anti-Rabbit (Agilent Dako) for 45 min at room temperature (human tissues), or ii ) a rabbit polyclonal anti-GSTO1 (LSBio Medical; 1:2000) overnight at 4°C and MACH 2 Rabbit HRP-Polymer for 30 min (Biocare Medical) at room temperature (mouse tissues).

Techniques: Expressing, RNA Sequencing, Generated, Gene Expression

Journal: Cancer letters

Article Title: Protective role of epithelial deoxyhypusine synthase in colorectal carcinogenesis caused by deletion of the adenomatous polyposis coli gene

doi: 10.1016/j.canlet.2025.218002

Figure Lengend Snippet: Apc fl/fl ( A ) and Apc fl/fl ;Dhps fl/fl ( A;D ) mice were treated or not with TAM. After 35 days, mice were euthanized, and the colon was removed. In TAM-treated mice, non-tumor (NT) and tumor (T) tissues were isolated. DNA was extracted from the tissues and the presence of WT and deleted alleles of Apc and Dhps genes was assessed by PCR ( A ); these gels are representative images of experiments performed on 3 mice per group. Total RNA was extracted from biopsies and the level of Apc and Dhps mRNA was determined by RT-real-time PCR ( B ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by ANOVA and Tukey test.

Article Snippet: Slides were then sequentially incubated with i ) the rabbit polyclonal anti-DHPS Ab (Proteintech; 1:1000) overnight at 4°C and EnVision+/HRP Labelled Polymer Anti-Rabbit (Agilent Dako) for 45 min at room temperature (human tissues), or ii ) a rabbit polyclonal anti-GSTO1 (LSBio Medical; 1:2000) overnight at 4°C and MACH 2 Rabbit HRP-Polymer for 30 min (Biocare Medical) at room temperature (mouse tissues).

Techniques: Isolation, Real-time Polymerase Chain Reaction

Journal: Cancer letters

Article Title: Protective role of epithelial deoxyhypusine synthase in colorectal carcinogenesis caused by deletion of the adenomatous polyposis coli gene

doi: 10.1016/j.canlet.2025.218002

Figure Lengend Snippet: Apc fl/fl ( A ; n = 29) and Apc fl/fl ;Dhps fl/fl mice ( A;D ; n = 22) were given TAM; there were also 7 Apc fl/fl and 6 Apc fl/fl ;Dhps fl/fl sham-treated animals. Survival ( A ) and body weights ( B ) were monitored for 35 days. At sacrifice, colons were harvested, opened longitudinally and the number of mice with visible tumors was determined ( C ). Tumors were counted ( D ) and measured ( E ) to calculate the total tumor burden per colon ( F ). Note that there were no tumors in animals that did not receive TAM. H&E staining ( G ; scale bars, 50 μm) was used to assess the number of histologic adenomas ( H ) and the grade of dysplasia ( I ). In panel F , we show a small adenoma circled by a dotted line and LGD at higher magnification in the Apc fl/fl + TAM group; a larger tumor and HGD is shown for the Apc fl/fl ;Dhps fl/fl + TAM group. In A , P was calculated by Log-rank (Mantel-Cox) test. In B , * P < 0.05 compared to Apc fl/fl ; §§§ P < 0.001 and §§§§ P < 0.0001 versus Apc fl/fl ;Dhps fl/fl ; # # P < 0.01 compared to Apc fl/fl + TAM by two-way ANOVA and Tukey test. P was determined by Fisher’s exact test for C and by Chi-square for E and I . In panels D , F , and H , ** P < 0.01, *** P < 0.001 by Student’s t test.

Article Snippet: Slides were then sequentially incubated with i ) the rabbit polyclonal anti-DHPS Ab (Proteintech; 1:1000) overnight at 4°C and EnVision+/HRP Labelled Polymer Anti-Rabbit (Agilent Dako) for 45 min at room temperature (human tissues), or ii ) a rabbit polyclonal anti-GSTO1 (LSBio Medical; 1:2000) overnight at 4°C and MACH 2 Rabbit HRP-Polymer for 30 min (Biocare Medical) at room temperature (mouse tissues).

Techniques: Staining

Journal: Cancer letters

Article Title: Protective role of epithelial deoxyhypusine synthase in colorectal carcinogenesis caused by deletion of the adenomatous polyposis coli gene

doi: 10.1016/j.canlet.2025.218002

Figure Lengend Snippet: ( A ) A label-free quantitative proteomic analysis was used to determine the differential expression of the proteins isolated from non-tumor and tumor tissues of Apc fl/fl and Apc fl/fl ;Dhps fl/fl mice treated with TAM compared to sham-treated mice ( n = 5 mice per group). The complete list of proteins identified in the analysis is provided in Supplementary Table 2 . The Venn diagrams summarize the number of proteins significantly upregulated or downregulated in each comparison ( A ); the names of the proteins involved in aldehyde detoxification and significantly affected are provided. ( B ) IHC for GSTO1 in the colon of Apc fl/fl and Apc fl/fl ;Dhps fl/fl mice ± TAM. These photomicrographs are representative of 3 sham-treated mice per genotype and 5–7 TAM-treated animals per genotype; scale bars, 50 μm. ( C ) The quantification of the IHC for GSTO1 in the tumors, performed in a blinded manner by our GI pathologist (M.B.P). ( D ) Quantification by LC-MS/MS of MDA-dilysyl crosslinks in tumors (T) or non-tumor (NT) tissues of Apc fl/fl and Apc fl/fl ;Dhps fl/fl mice treated or not with TAM. * P < 0.05 and ** P < 0.01 by Student’s t test ( C ) or ANOVA and Šídák’s multiple comparisons test ( D ).

Article Snippet: Slides were then sequentially incubated with i ) the rabbit polyclonal anti-DHPS Ab (Proteintech; 1:1000) overnight at 4°C and EnVision+/HRP Labelled Polymer Anti-Rabbit (Agilent Dako) for 45 min at room temperature (human tissues), or ii ) a rabbit polyclonal anti-GSTO1 (LSBio Medical; 1:2000) overnight at 4°C and MACH 2 Rabbit HRP-Polymer for 30 min (Biocare Medical) at room temperature (mouse tissues).

Techniques: Quantitative Proteomics, Isolation, Comparison, Liquid Chromatography with Mass Spectroscopy

Journal: Cancer letters

Article Title: Protective role of epithelial deoxyhypusine synthase in colorectal carcinogenesis caused by deletion of the adenomatous polyposis coli gene

doi: 10.1016/j.canlet.2025.218002

Figure Lengend Snippet: Apc fl/fl ;Dhps fl/fl mice were treated or not with TAM ± 2-HOBA. ( A ) Body weights were monitored weekly and are shown as percentage of initial body weight. ( B-D ) Tumor number ( B ), tumor size ( C ), and total tumor burden ( D ) were determined in the colon. ( E ) The H&E staining shows a large tumor with a complex growth pattern indicating HGD in the TAM group and one smaller tumor formed by densely packed neoplastic crypts with LGD for the TAM group in mice treated with 2-HOBA; scale bar, 100 μm for the mice without TAM and 50 μm for TAM-treated animals. ( F ) The grade of dysplasia was determined from the H&E staining. * P < 0.05, ** P < 0.01 and *** P < 0.001 by two-way ANOVA and Tukey test ( A ) and Student’s t test ( B , D ). For C and F , P was determined by Chi-square and Fisher’s exact test, respectively.

Article Snippet: Slides were then sequentially incubated with i ) the rabbit polyclonal anti-DHPS Ab (Proteintech; 1:1000) overnight at 4°C and EnVision+/HRP Labelled Polymer Anti-Rabbit (Agilent Dako) for 45 min at room temperature (human tissues), or ii ) a rabbit polyclonal anti-GSTO1 (LSBio Medical; 1:2000) overnight at 4°C and MACH 2 Rabbit HRP-Polymer for 30 min (Biocare Medical) at room temperature (mouse tissues).

Techniques: Staining

Journal: Cancer letters

Article Title: Protective role of epithelial deoxyhypusine synthase in colorectal carcinogenesis caused by deletion of the adenomatous polyposis coli gene

doi: 10.1016/j.canlet.2025.218002

Figure Lengend Snippet: ( A ) The subcellular localization of NRF2 in the colon tumors of Apc fl/fl and Apc fl/fl ;Dhps fl/fl mice, treated or not with TAM ± 2-HOBA, was assessed by immunofluorescence and the images are representatives of 4 animals per group; green, NRF2; blue, nucleus; turquoise, double staining. Scale bar, 100 μm. ( B ) The expression of Slc7a11 and Hmox1 was analyzed by RT-real-time PCR in tumor (T) and non-tumor tissues (NT) from Apc fl/fl ( A ) and Apc fl/fl ;Dhps fl/fl ( A;D ). ( C ) Gene expression in Apc fl/fl ;Dhps fl/fl ± TAM ± 2-HOBA. In B and C, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 by ANOVA and Šídák’s multiple comparisons test.

Article Snippet: Slides were then sequentially incubated with i ) the rabbit polyclonal anti-DHPS Ab (Proteintech; 1:1000) overnight at 4°C and EnVision+/HRP Labelled Polymer Anti-Rabbit (Agilent Dako) for 45 min at room temperature (human tissues), or ii ) a rabbit polyclonal anti-GSTO1 (LSBio Medical; 1:2000) overnight at 4°C and MACH 2 Rabbit HRP-Polymer for 30 min (Biocare Medical) at room temperature (mouse tissues).

Techniques: Activation Assay, Immunofluorescence, Double Staining, Expressing, Real-time Polymerase Chain Reaction, Gene Expression

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Schematic illustration of SARS-CoV-2 spike protein and ACE2 receptor binding. (a) SARS-CoV-2 binding to the ACE2 receptor on the host cell surface. (b) Binding of ACE2 and spike protein along with illustration of the spike protein subunits, S1 and S2. (c) Schematic showing the distribution of the spike protein in solution. (d) Schematic showing the distribution of ACE2 in suspension. (e) Distribution of ACE2 and S protein after binding in solution.

Article Snippet: The SARS-CoV-2 S1 (ProSci, Cat.No.97-087) and S2 (ProSci Cat.No.97-079) recombinant proteins were purchased in lyophilized form and resuspended according to the manufacturer’s instructions at a stock concentration of 600 µg/ml for S1 and 200 µg/ml for S2 protein in sterile water.

Techniques: Binding Assay

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Optical measurements of the spike protein subunits S1 and S2: (a) Measured responses for spikes proteins S1 and S2 at the highest concentration individually (S1B and S2B, respectively), along with their corresponding blanks. (b) Time domain measurements of the microcentrifuge tube, the blank (shown in gray circles) versus water (red circles) at a wavelength of 623 nm. (c) Measured optical responses for the mixed protein samples versus time. Samples S1B and S2B were at 5000 copies per ml, S2C, S2D, S2E and S2F are the serial dilutions of S2B at 10-, 100-, 1,000- and 10,000-fold, respectively. (d) Relative change in light intensity per light path versus loaded mass. All optical responses were measured at 623 nm. Light intensity was measured as arbitrary units (a.u).

Article Snippet: The SARS-CoV-2 S1 (ProSci, Cat.No.97-087) and S2 (ProSci Cat.No.97-079) recombinant proteins were purchased in lyophilized form and resuspended according to the manufacturer’s instructions at a stock concentration of 600 µg/ml for S1 and 200 µg/ml for S2 protein in sterile water.

Techniques: Concentration Assay