Journal: Frontiers in Oncology

Article Title: Prognostic value and experimental validation of atherosclerosis-derived pathogenic genes in colorectal cancer

doi: 10.3389/fonc.2025.1728087

Figure Lengend Snippet: Validation of the expression of 6 hub genes in CRC and AS by molecular biology experiments. (A-F) RT-qPCR validation of the differential expression of prognostic genes between normal and tumor tissues in CRC patients. (G) Western blot validation of the differential expression of 2 newly identified genes (HMMR and PALB2) between normal and tumor tissues in CRC. (H) Quantitative analysis of the relative grayscale values of Western blot bands for the 2 newly identified genes (HMMR and PALB2) in normal and tumor tissues of CRC. Data are presented as mean ± standard deviation (SD), with statistical significance indicated as * p < 0.05, *** p < 0.001, and **** p < 0.0001. ns indicates no statistical significance.

Article Snippet: The membrane was blocked with QuickBlockTM Blocking Buffer for Western Blot (Beyotime Biotechnology, Jiangsu, China) at room temperature for 1 h, washed with TBST, and incubated overnight at 4°C with primary antibodies: HMMR (1:2000, 15820-1-AP, Proteintech), PALB2 (1:2000, 14340-1-AP, Proteintech), and β-actin (1:10,000, 66009-1-Ig, Proteintech).

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Quantitative Proteomics, Western Blot, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Prognostic value and experimental validation of atherosclerosis-derived pathogenic genes in colorectal cancer

doi: 10.3389/fonc.2025.1728087

Figure Lengend Snippet: IHC staining validation. (A) The differential expression of PALB2 and HMMR in CRC tissues compared with normal tissues; (B) displays the differential expression of HMMR , PALB2 and PRR11 in AS tissues versus normal tissues. * p < 0.05.

Article Snippet: The membrane was blocked with QuickBlockTM Blocking Buffer for Western Blot (Beyotime Biotechnology, Jiangsu, China) at room temperature for 1 h, washed with TBST, and incubated overnight at 4°C with primary antibodies: HMMR (1:2000, 15820-1-AP, Proteintech), PALB2 (1:2000, 14340-1-AP, Proteintech), and β-actin (1:10,000, 66009-1-Ig, Proteintech).

Techniques: Immunohistochemistry, Biomarker Discovery, Quantitative Proteomics

Journal: DNA Repair

Article Title: Specific killing of DNA damage-response deficient cells with inhibitors of poly(ADP-ribose) glycohydrolase

doi: 10.1016/j.dnarep.2017.02.010

Figure Lengend Snippet: Summary of results following depletion of target DDR gene using siRNA in combination with PARG siRNA or PARG inhibitors.

Article Snippet: This membrane was immunoblotted with antibodies against Poly(ADP-ribose) (1:400, 10H Enzo Life Sciences), PARG (1:250, Abcam), PARP1 (1:1000, Santa Cruz), BRCA1 (1:500, Santa Cruz), BARD1 (1:500, H-300, Santa Cruz), PTEN (1:1000, Cell Signaling), PALB2 (1:500, Novus Biologicals), FAM175A (1:1000, Bethyl) and β-tubulin (1:2000, Sigma) each diluted in 5% milk and incubated at 4 °C overnight.

Techniques:

Journal: Human Genetics and Genomics Advances

Article Title: Prediction of breast cancer risk based on flow variant analysis of circulating peripheral blood mononuclear cells

doi: 10.1016/j.xhgg.2022.100085

Figure Lengend Snippet: Expression plasmid rescue of genetic variants in LCLs (A–D) Gene rescue for (A) BRCA1 , (B) BRCA2 , (C) ATM , and (D) PALB2 variants by RCS by boxplots. P-values for pairwise comparisons are shown.

Article Snippet: Expression plasmids (1 μg) for BRCA1 (pDEST-FRT/T0-GFP-BRCA1, cat. no. 71116, GFP tag), BRCA2 (pMH-SFB-BRCA2, cat. no. 99395, SFP tag), PALB2 (pDEST-FRT/T0-GFP-PALB2, cat. no. 71113, GFP tag), and ATM (pcDNA3.1(+)FLAG-His-ATM WT, cat. no. 31985, Addgene, Watertown, MA) were transfected into WT LCLs or those with P/LP or B/LB variants.

Techniques: Expressing, Plasmid Preparation

Journal: BMC cancer

Article Title: NUSAP1 promotes pancreatic ductal adenocarcinoma progression by drives the epithelial-mesenchymal transition and reduces AMPK phosphorylation.

doi: 10.1186/s12885-024-11842-5

Figure Lengend Snippet: Fig. 6 Decreasing NUSAP1 expression inhibits PDAC cell proliferation. a Relative expression levels of NUSAP1 among the PDAC cell lines. b mRNA expression levels of NUSAP1 in PANC-1 and CAPAN-1 cells were detected by reverse transcription quantitative PCR after transfection of si-NUSAP1. c The protein expression level of NUSAP1 decreased significantly after transfection of cells with si-NUSAP1. d The proliferation of PDAC cells was measured using CCK-8 assays. e Visualization of DNA replication by EdU incorporation. Cell nuclei stained red represent DNA replication. f A significant reduction in colony formation ability was observed after transfection with si-NUSAP1. g Apoptosis rates of the si-NUSAP1 cell lines. Student’s t test was used for comparisons between groups. All experiments were performed in triplicate. *P < 0.05. NC, negative control; si, small interfering RNA.

Article Snippet: The siRNA targeting NUSAP1 and scrambled negative control siRNA (si-NC) were purchased from Santa Cruz Biotechnology, Inc. (cat. nos. sc-93,396 and sc-37,007).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Transfection, CCK-8 Assay, Staining, Negative Control, Small Interfering RNA

Journal: BMC cancer

Article Title: NUSAP1 promotes pancreatic ductal adenocarcinoma progression by drives the epithelial-mesenchymal transition and reduces AMPK phosphorylation.

doi: 10.1186/s12885-024-11842-5

Figure Lengend Snippet: Fig. 7 Decreasing the expression of NUSAP1 inhibits the migration and invasion of PDAC cells. a Migration of PANC-1 cells in the si-NC and si-NUSAP1 groups according to Wound healing assays. b Migration of PANC-1 and CAPAN-1 cells in the si-NC and si-NUSAP1 groups according to Transwell assays. c Invasion of PANC-1 and CAPAN-1 cells in the si-NC and si-NUSAP1 groups according to Transwell assays. Student’s t test was used for comparisons between groups. All experiments were performed in triplicate. *P < 0.05. NC, negative control; si, small interfering RNA.

Article Snippet: The siRNA targeting NUSAP1 and scrambled negative control siRNA (si-NC) were purchased from Santa Cruz Biotechnology, Inc. (cat. nos. sc-93,396 and sc-37,007).

Techniques: Expressing, Migration, Negative Control, Small Interfering RNA

Journal: BMC cancer

Article Title: NUSAP1 promotes pancreatic ductal adenocarcinoma progression by drives the epithelial-mesenchymal transition and reduces AMPK phosphorylation.

doi: 10.1186/s12885-024-11842-5

Figure Lengend Snippet: Fig. 9 NUSAP1 promoted progression of PDAC in vivo. a We randomly divided the mice into the NC and the KD groups (n = 5), PANC-1 cells stably transfected with an empty vector or the NUSAP1 shRNA were subcutaneously inoculated in them respectively. Then we treated the mice as described in the Methods. b The tumor sizes were tested using Vernier calipers. Tumor growth curves were constructed based on the tumor volumes measured in the mice. c Quantification of the average weights of collected tumors from the above experiments. d The expression of NUSAP1, Ki-67, BAX and Cleaved caspase-3 were determined in tumor tissue sections from the xenografts using IHC (scale bar, 40 μm, n = 5). e The expression of NUSAP1, Ki-67, BAX, and Cleaved caspase-3 in tumor tissue between the NC and the KD groups, as determined by the IHC score. Student’s t test was used for comparisons between groups. *P < 0.05, **P < 0.01. NC, negative control; KD, knockdown

Article Snippet: The siRNA targeting NUSAP1 and scrambled negative control siRNA (si-NC) were purchased from Santa Cruz Biotechnology, Inc. (cat. nos. sc-93,396 and sc-37,007).

Techniques: In Vivo, Stable Transfection, Transfection, Plasmid Preparation, shRNA, Construct, Expressing, Negative Control, Knockdown