npc2  (Proteintech)

Journal: Journal of Inflammation Research

Article Title: Integrative Machine Learning Analysis of Programmed Cell Death Pathways Identifies Novel Diagnostic Biomarkers for Atrial Fibrillation

doi: 10.2147/JIR.S568171

Figure Lengend Snippet: Functional annotation and immune landscape of PCDscore. ( A ) Distribution of PCDscores of AF patients; ( B ) The KEGG functional enrichment of differential genes between high and low risk groups; ( C ) Heatmap illustrating the immune landscape between the high and low risk groups; ( D ) Heatmap illustrating the immune modulators between the two high and low risk group; ( E ) Correlation matrix between PCDscore, NPC2, SGPL1 and various immune cell types.

Article Snippet: Membranes were blocked with 5% non-fat milk (TBST, 1 h, 20–25 °C), incubated with primary antibodies against NPC2 and PTGDS (Proteintech, Wuhan, China) and against RCAN1 and SGPL1 (Cell Signaling Technology, MA, USA) overnight (4 °C), washed, and probed with HRP-conjugated secondaries (1 h, 20–25 °C).

Techniques: Functional Assay

Journal: Journal of Inflammation Research

Article Title: Integrative Machine Learning Analysis of Programmed Cell Death Pathways Identifies Novel Diagnostic Biomarkers for Atrial Fibrillation

doi: 10.2147/JIR.S568171

Figure Lengend Snippet: Prediction of drugs and molecular docking. ( A ) The top 10 candidate drugs were predicted for hub genes using CMAP database. ( B – E ) Todralazine docking to NPC2 (PDB 5KWY), PTGDS (3O22), RCAN1 (6UUQ), and SGPL1 (8AYF) using CB-Dock2. Reported Vina scores indicate favorable binding (<−5).

Article Snippet: Membranes were blocked with 5% non-fat milk (TBST, 1 h, 20–25 °C), incubated with primary antibodies against NPC2 and PTGDS (Proteintech, Wuhan, China) and against RCAN1 and SGPL1 (Cell Signaling Technology, MA, USA) overnight (4 °C), washed, and probed with HRP-conjugated secondaries (1 h, 20–25 °C).

Techniques: Binding Assay

Journal: Journal of Inflammation Research

Article Title: Integrative Machine Learning Analysis of Programmed Cell Death Pathways Identifies Novel Diagnostic Biomarkers for Atrial Fibrillation

doi: 10.2147/JIR.S568171

Figure Lengend Snippet: Experimental validation in HL-1 cells and human PBMCs. qRT-PCR analysis of NPC2 ( A ), RCAN1 ( B ), SGPL1 ( C ) and PTGDS ( D ) expression in AF cell models; qRT-PCR analysis of NPC2 ( E ) and SGPL1 ( F ) in PBMCs from AF patients and healthy controls. ( G ) Western blot analysis of NPC2, RCAN1, SGPL1 and PTGDS expression in AF cell models. * P <0.05; ** P <0.01.

Article Snippet: Membranes were blocked with 5% non-fat milk (TBST, 1 h, 20–25 °C), incubated with primary antibodies against NPC2 and PTGDS (Proteintech, Wuhan, China) and against RCAN1 and SGPL1 (Cell Signaling Technology, MA, USA) overnight (4 °C), washed, and probed with HRP-conjugated secondaries (1 h, 20–25 °C).

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

Journal: Functional & Integrative Genomics

Article Title: ALKBH5-mediated NPC2 mRNA m 6 A demethylation promotes resistance to oxaliplatin in colorectal cancer

doi: 10.1007/s10142-025-01651-9

Figure Lengend Snippet: ALKBH5 increases NPC2 expression in the oxaliplatin resistant CRC. ( A , B ) HCT116 and LoVo cells were treated with oxaliplatin (5 µg/mL) for 48 h, then the mRNA level of m 6 A regulators (METTL3, METTL14, METTL16, WATP, ALKBH5, FTO, IGF2BP1, IGF2BP2, IGF2BP3, YTHDF1, YTHDF2, YTHDF3, YTHDC1, YTHDC2) were measured by RT-qPCR. ( C ) HCT116 cells were treated with oxaliplatin (5 µg/mL) for 48 h, the expression of NPC2 mRNA was measured by RT-qPCR. ( D ) HCT116 cells were treated with oxaliplatin (5 µg/mL) for 48 h, the protein expression of NPC2 and ALKBH5 was measured by Western blot analysis. ( E ) HCT116 cells were transfected with a luciferase reporter construct containing the ALKBH5 promoter, followed by treatment with oxaliplatin (5 µg/mL) for 48 h. Luciferase activity was measured and normalized to Renilla. ( F , G ) HCT116 cells were transfected with ALKBH5 plasmid, then the expression level of ALKBH5 and NPC2 were measured by RT-qPCR ( F ) and Western blot analysis ( G ). ( H ) HCT116 cells were transfected with ALKBH5 plasmid, and then treated with oxaliplatin (2 µg/mL), the proliferation of cells was determined by the colony formation assay. ( I ) Counting and statistical analysis of the colony numbers. ( J , K ) HCT116 cells were transfected with NC or ALKBH5 siRNA, then the expression level of ALKBH5 and NPC2 were measured by RT-qPCR ( J ) and Western blot analysis ( K ). ( L ) HCT116-OXR cells were transfected with control or ALKBH5-siRNA, and then treated with oxaliplatin (2 µg/mL), the proliferation of cells was determined by the colony formation assay. ( M ) Counting and statistical analysis of the colony numbers. ( N ) HCT116 cells were transfected with ALKBH5 plasmid, the m 6 A modifications of NPC2 mRNA were measured by MeRIP-qPCR. ( O ) HCT116 cells were transfected with ALKBH5 siRNA, the m 6 A modifications of NPC2 mRNA were measured by MeRIP-qPCR. ( P ) HCT116-OXR cells were treated with oxaliplatin (5 µg/mL), and RIP-PCR was performed to detect the molecular interaction within ALKBH5 and NPC2. ( Q ) The binding capacity between NPC2 mRNA and ALKBH5 protein or YTHDF2 protein in HCT116 cells were examined by RNA pulldown and western blot analysis. ( R ) HCT116 cells were transfected with ALKBH5 plasmid or ALKBH5 siRNA, then treated with ActD (5 µg/mL) for the indicated times. The expression of NPC2 mRNA was examined with RT-qPCR. Data are presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001)

Article Snippet: The siRNAs targeting NPC2, METTL3, FTO, ALKBH5 and YTHDF2 were purchased from GenePharma.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Luciferase, Construct, Activity Assay, Plasmid Preparation, Colony Assay, Control, Binding Assay

Journal: Functional & Integrative Genomics

Article Title: ALKBH5-mediated NPC2 mRNA m 6 A demethylation promotes resistance to oxaliplatin in colorectal cancer

doi: 10.1007/s10142-025-01651-9

Figure Lengend Snippet: YTHDF2 promotes the degradation of NPC2 mRNA in the oxaliplatin resistant CRC. ( A - E ) HCT116 cells were transfected with siRNA of YTHDF1 ( A ), YTHDF2 ( B ), YTHDF3 ( C ), YTHDC1 ( D ), YTHDC2 ( E ), then the mRNA level of YTH domain-containing proteins and NPC2 were measured by RT-qPCR. ( F ) HCT116 cells were transfected with NC or YTHDF2 siRNA, then the protein level of YTHDF2 and NPC2 were measured by Western blot analysis. ( G ) HCT116 cells were transfected with control or YTHDF2-siRNA, and then treated with oxaliplatin (2 µg/mL), the proliferation of cells was determined by the colony formation assay. ( H ) Counting and statistical analysis of the colony numbers. ( I ) HCT116-OXR cells were treated with oxaliplatin (5 µg/mL), and RIP-PCR was performed to detect the molecular interaction within YTHDF2 and NPC2. ( J ) HCT116 cells were transfected with NC or YTHDF2 siRNA, then treated with ActD (5 µg/mL) for the indicated times. The expression of NPC2 mRNA was examined with RT-qPCR. ( K ) HCT116 cells were transfected with NC or YTHDF2-siRNA then with or without ALKBH5 knockdown. The protein level of ALKBH5, YTHDF2 and NPC2 were measured by Western blot analysis. ( L ) HCT116 cells were transfected with Vector or YTHDF2 plasmid then with or without ALKBH5 overexpression. The protein level of ALKBH5, YTHDF2 and NPC2 were measured by Western blot analysis. Data are presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001)

Article Snippet: The siRNAs targeting NPC2, METTL3, FTO, ALKBH5 and YTHDF2 were purchased from GenePharma.

Techniques: Transfection, Quantitative RT-PCR, Western Blot, Control, Colony Assay, Expressing, Knockdown, Plasmid Preparation, Over Expression