rabbit anti hypoxanthine phosphoribosyltransferase 1  (Proteintech)

Journal: Molecular Cancer Research

Article Title: Metabolic Hallmarks for Purine Nucleotide Biosynthesis in Small Cell Lung Carcinoma

doi: 10.1158/1541-7786.MCR-23-0386

Figure Lengend Snippet: HPRT1 expression in SCLC tissues and cell lines. A and B, IHC staining of HPRT1 in tumors of SCLC patients ( n = 59) with HPRT1-positive (HPRT+, A ) or HPRT-negative (HPRT−, B ) expression. Scale bar = 50 μmol/L. C, Levels HPRT1 protein expression in the 24 SCLC cell lines. GAPDH was used as a housekeeping protein.

Article Snippet: Primary antibodies were diluted in Antibody Diluent (Agilent) at a 1:100 dilution and incubated overnight at 4°C with a rabbit mAb against HPRT1 (ab109021; Abcam).

Techniques: Expressing, Immunohistochemistry

Journal: Molecular Cancer Research

Article Title: Metabolic Hallmarks for Purine Nucleotide Biosynthesis in Small Cell Lung Carcinoma

doi: 10.1158/1541-7786.MCR-23-0386

Figure Lengend Snippet: Loss of HPRT1 leads to antifolate sensitivity in SCLC cells. A–C, HPRT1 protein expression in DMS273 cells ( A ), H1048 cells ( B ), and SBC-5 cells ( C ) and their HPRT1 -knockout (KO) clones (DH1–5). Two clones (indicated in red) from each SCLC cell line were used for further analyses. D–F, Effect of HPRT1 KO on cell survival in DMS273 ( D ), H1048 ( E ), and SBC-5 ( F ) cells under lemetrexol (LMX) treatment conditions for 72 hours. Gray lines are parent cells, and red and blue lines are HPRT1-deleted cells ( n = 6–8). G–I, Effect of ectopic HPRT1 expression on cell survival in HPRT1 -KO cells treated with LMX for 72 hours. DMS273/ HPRT1 -KO ( G ), H1048/ HPRT1 -KO ( H ), and SBC-5/HPRT1-KO cells ( I ). Red lines, control vector; green lines, HPRT1 expression. J, Effect of 100 μmol/L hypoxanthine (HXN) on HPRT1 -KO DMS273 cells transfected with the control vector or HPRT1 cDNA. Data are presented as the mean ± SD ( n = 6–9). Two-tailed Student t test and P values are indicated as *, P < 0.05, **, P < 0.01, and ***, P < 0.005.

Article Snippet: Primary antibodies were diluted in Antibody Diluent (Agilent) at a 1:100 dilution and incubated overnight at 4°C with a rabbit mAb against HPRT1 (ab109021; Abcam).

Techniques: Expressing, Knock-Out, Clone Assay, Plasmid Preparation, Transfection, Two Tailed Test

Journal: Molecular Cancer Research

Article Title: Metabolic Hallmarks for Purine Nucleotide Biosynthesis in Small Cell Lung Carcinoma

doi: 10.1158/1541-7786.MCR-23-0386

Figure Lengend Snippet: Effect of HPRT1 on tumor growth in vivo . A–C, Growth of HPRT1-deleted tumors. Tumor size (mm 3 ) was measured over time in vivo in immunodeficient mice ( n = 3–8) xenografted with DMS273 ( A ), H1048 ( B ), and SBC-5 ( C ) cell lines. Wild-type (a, gray line) and HPRT1-deleted (red or blue line) cells are shown. D, Tumor growth in the HPRT1 rescue tumors. HPRT1-deleted DMS273 cells transfected with either the vector (b, red line) or HPRT1 cDNA (b, green line) are shown. Data are presented as the mean ± SD. Two-tailed Student t test and P values are indicated as *, P < 0.05 and ***, P < 0.005.

Article Snippet: Primary antibodies were diluted in Antibody Diluent (Agilent) at a 1:100 dilution and incubated overnight at 4°C with a rabbit mAb against HPRT1 (ab109021; Abcam).

Techniques: In Vivo, Transfection, Plasmid Preparation, Two Tailed Test

Journal: Molecular Cancer Research

Article Title: Metabolic Hallmarks for Purine Nucleotide Biosynthesis in Small Cell Lung Carcinoma

doi: 10.1158/1541-7786.MCR-23-0386

Figure Lengend Snippet: Metabolic alterations in HPRT1 -knockout tumors. A, Volcano plots showing differences in metabolite levels in tumors from HPRT1-deleted DMS273 cells. Metabolomic analysis was conducted on SCLC tumors ( n = 5) using CE/MS. B, Metabolite pathway enrichment analysis of metabolite differences between wild-type and HPRT-knockout (KO) tumors. C, Levels of AICAR and metabolites in the pentose phosphate pathway in tumors from wild-type and HPRT1 -KO DMS273 cells. D, Rescue effect of HPRT1 in tumors from HPRT1 -KO DMS273 cells transfected with the control vector (KO+V) and HPRT1 cDNA (KO+H). Data are presented as the mean ± SD. Two-tailed Student t test and P values are indicated as *, P < 0.05 and ***, P < 0.005.

Article Snippet: Primary antibodies were diluted in Antibody Diluent (Agilent) at a 1:100 dilution and incubated overnight at 4°C with a rabbit mAb against HPRT1 (ab109021; Abcam).

Techniques: Knock-Out, Transfection, Plasmid Preparation, Two Tailed Test

Journal: Molecular Cancer Research

Article Title: Metabolic Hallmarks for Purine Nucleotide Biosynthesis in Small Cell Lung Carcinoma

doi: 10.1158/1541-7786.MCR-23-0386

Figure Lengend Snippet: Metabolic flux analysis for purine nucleotide biosynthesis in DMS273 cells with or without HPRT1 expression. A, Schematic describing the experimental design used to analyze the metabolite incorporation of [ 13 C 2 , 15 N]-glycine (M+3) in the purine de novo pathway and [ 13 C 5 , 15 N 4 ]-hypoxanthine (M+9) in the purine salvage pathway. B, Intracellular levels of Gly (M+3), AMP (M+3), ADP (M+3), and ATP (M+3) derived from the purine de novo pathway in the HPRT1 -knockout (KO) DMS273 cells transfected with control vector (KO+V) or HPRT1 cDNA (KO+H). The cells were treated with [ 13 C 2 , 15 N]-glycine (M+3) and [ 13 C 5 , 15 N 4 ]-hypoxanthine (M+9) for 0, 2, and 6 hours. C, Intracellular levels of hypoxanthine (M+9), AMP (M+9), ADP (M+9), and ATP (M+9) derived from the purine salvage pathway in the KO+V or KO+H cells. D, Effect of LMX on tumors from KO+V or KO+H cells ( n = 8). After the tumor size reached >100 mm 3 , lemetrexol (25 mg/kg) was administered intravenously every 2 days. Data are presented as mean ± SD. Two-tailed Student t test and P values are indicated as *, P < 0.05.

Article Snippet: Primary antibodies were diluted in Antibody Diluent (Agilent) at a 1:100 dilution and incubated overnight at 4°C with a rabbit mAb against HPRT1 (ab109021; Abcam).

Techniques: Expressing, Derivative Assay, Knock-Out, Transfection, Plasmid Preparation, Two Tailed Test

Journal: eLife

Article Title: SIRT2 inhibition protects against cardiac hypertrophy and ischemic injury

doi: 10.7554/eLife.85571

Figure Lengend Snippet: Primer sequences.

Article Snippet: Antibody , Rabbit polyclonal anti-HPRT antibody , Proteintech , 150-59-1-AP , WB (1:5000).

Techniques:

Journal: eLife

Article Title: SIRT2 inhibition protects against cardiac hypertrophy and ischemic injury

doi: 10.7554/eLife.85571

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal anti-HPRT antibody , Proteintech , 150-59-1-AP , WB (1:5000).

Techniques: Knock-Out, Activity Assay, In Vivo, Protease Inhibitor, Transfection, SYBR Green Assay, cDNA Synthesis, Bicinchoninic Acid Protein Assay, Extraction, Staining, Sequencing, Recombinant, Plasmid Preparation, Software

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: HPRT1 is upregulated and predicts a poor prognosis in HNSCC. (A, B) Expression of HPRT1 in both unpaired and paired HNSCC and normal tissues were evaluated using TCGA data. ( C ) qRT-PCR analysis of the expression of HPRT1 mRNA in HNSCC and para-cancer tissues. ( D ) TCGA data indicating the prognostic values of HPRT1 in HNSCC, including overall survival (OS), disease-specific survival (DSS), and progression-free interval (PFI). ( E ) Representative images of immunohistochemical (IHC) staining for HPRT1 protein in HNSCC and para-cancer tissues. *** P < 0.001.

Article Snippet: The used primary antibodies were rabbit anti-HPRT1 (1:200, BBI Life Sciences Corp., Shanghai, China), mouse anti-Tubulin (1:1000, Proteintech, Wuhan, China), and HRP-conjugated goat anti-mouse/rabbit IgG (1:3000, BBI Life Sciences Corp., Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemistry

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: The correlation between HPRT1 expression and the clinicopathological parameters of patients with HNSCC (n = 60, Chi-Squared Test).

Article Snippet: The used primary antibodies were rabbit anti-HPRT1 (1:200, BBI Life Sciences Corp., Shanghai, China), mouse anti-Tubulin (1:1000, Proteintech, Wuhan, China), and HRP-conjugated goat anti-mouse/rabbit IgG (1:3000, BBI Life Sciences Corp., Shanghai, China).

Techniques: Expressing

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: HPRT1 depletion suppresses proliferation and invasion of HNSCC cells in vitro . ( A ) Western blot analysis of HPRT1in HNSCC cells including HOK, FaDu, SCC-4, and CAL-27 cells. ( B ) Western blot analysis of HPRT1 in HNSCC cells infected with shHPRT1 lentivirus particles. ( C ) CCK-8, ( D ) plate clone formation, and ( E ) transwell invasion assays indicating the cell viability, proliferation, and invasion of HNSCC cells with HPRT1 depletion. ( F ) Western blot analysis of MMP2 in HNSCC cells with HPRT1 knockdown. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The used primary antibodies were rabbit anti-HPRT1 (1:200, BBI Life Sciences Corp., Shanghai, China), mouse anti-Tubulin (1:1000, Proteintech, Wuhan, China), and HRP-conjugated goat anti-mouse/rabbit IgG (1:3000, BBI Life Sciences Corp., Shanghai, China).

Techniques: In Vitro, Western Blot, Infection, CCK-8 Assay

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: HPRT1 depletion suppresses the proliferation of HNSCC cells in vivo . （A）Images of nude mice and xenograft tumors derived from HNSCC cells with HPRT1 depletion. ( B ) Volume curve of xenograft tumors. ( C ) Weight of xenograft tumors at the endpoint. (D) IHC staining of HPRT1 and Ki-67 in xenograft tumors. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The used primary antibodies were rabbit anti-HPRT1 (1:200, BBI Life Sciences Corp., Shanghai, China), mouse anti-Tubulin (1:1000, Proteintech, Wuhan, China), and HRP-conjugated goat anti-mouse/rabbit IgG (1:3000, BBI Life Sciences Corp., Shanghai, China).

Techniques: In Vivo, Derivative Assay, Immunohistochemistry

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: MiR-125b-5p targets HPRT1 in HNSCC. ( A ) qRT-PCR results analysis of HPRT1 hnRNA in HOK and HNSCC cells. ( B ) qRT-PCR and ( C ) Western blot analyses of HPRT1 in FaDu and SCC-4 cells transfected with miR-125b-5p mimic. ( D ) Pearson correlation analysis between miR-125b-5p and HPRT1 expression in HNSCC tissues. ( E ) Dual-luciferase reporting assay was used to detect the relative luciferase intensity of FaDu and SCC-4 cells co-transfected with miR-125b-5p mimic and wild-type/mutant HPRT1. ** P < 0.01, *** P < 0.001.

Article Snippet: The used primary antibodies were rabbit anti-HPRT1 (1:200, BBI Life Sciences Corp., Shanghai, China), mouse anti-Tubulin (1:1000, Proteintech, Wuhan, China), and HRP-conjugated goat anti-mouse/rabbit IgG (1:3000, BBI Life Sciences Corp., Shanghai, China).

Techniques: Quantitative RT-PCR, Western Blot, Transfection, Expressing, Luciferase, Mutagenesis

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: miR-125b-5p inhibits HNSCC cell proliferation and invasion by targeting HPRT1. ( A ) Western blot analysis of HPRT1 and MMP2 in HNSCC cells co-transfected with HPRT1 and miR-125b-5p. ( B ) CCK-8, ( C ) plate clone formation, and ( D ) transwell invasion assays were used to evaluate cell viability, proliferation, and invasion of HNSCC cells co-transfected with HPRT1 and miR-125b-5p. * P < 0.05, ** P < 0.01, *** P < 0.001. # P < 0.05, ## P < 0.01.

Article Snippet: The used primary antibodies were rabbit anti-HPRT1 (1:200, BBI Life Sciences Corp., Shanghai, China), mouse anti-Tubulin (1:1000, Proteintech, Wuhan, China), and HRP-conjugated goat anti-mouse/rabbit IgG (1:3000, BBI Life Sciences Corp., Shanghai, China).

Techniques: Western Blot, Transfection, CCK-8 Assay

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: HPRT1 is upregulated and predicts a poor prognosis in HNSCC. (A, B) Expression of HPRT1 in both unpaired and paired HNSCC and normal tissues were evaluated using TCGA data. ( C ) qRT-PCR analysis of the expression of HPRT1 mRNA in HNSCC and para-cancer tissues. ( D ) TCGA data indicating the prognostic values of HPRT1 in HNSCC, including overall survival (OS), disease-specific survival (DSS), and progression-free interval (PFI). ( E ) Representative images of immunohistochemical (IHC) staining for HPRT1 protein in HNSCC and para-cancer tissues. *** P < 0.001.

Article Snippet: Then, slides were incubated overnight at 4 °C with the rabbit anti-HPRT1 antibody (dilution:1:50, BBI, Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining, Immunohistochemistry

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: The correlation between HPRT1 expression and the clinicopathological parameters of patients with HNSCC (n = 60, Chi-Squared Test).

Article Snippet: Then, slides were incubated overnight at 4 °C with the rabbit anti-HPRT1 antibody (dilution:1:50, BBI, Shanghai, China).

Techniques: Expressing

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: HPRT1 depletion suppresses proliferation and invasion of HNSCC cells in vitro . ( A ) Western blot analysis of HPRT1in HNSCC cells including HOK, FaDu, SCC-4, and CAL-27 cells. ( B ) Western blot analysis of HPRT1 in HNSCC cells infected with shHPRT1 lentivirus particles. ( C ) CCK-8, ( D ) plate clone formation, and ( E ) transwell invasion assays indicating the cell viability, proliferation, and invasion of HNSCC cells with HPRT1 depletion. ( F ) Western blot analysis of MMP2 in HNSCC cells with HPRT1 knockdown. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Then, slides were incubated overnight at 4 °C with the rabbit anti-HPRT1 antibody (dilution:1:50, BBI, Shanghai, China).

Techniques: In Vitro, Western Blot, Infection, CCK-8 Assay, Knockdown

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: HPRT1 depletion suppresses the proliferation of HNSCC cells in vivo . （A）Images of nude mice and xenograft tumors derived from HNSCC cells with HPRT1 depletion. ( B ) Volume curve of xenograft tumors. ( C ) Weight of xenograft tumors at the endpoint. (D) IHC staining of HPRT1 and Ki-67 in xenograft tumors. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Then, slides were incubated overnight at 4 °C with the rabbit anti-HPRT1 antibody (dilution:1:50, BBI, Shanghai, China).

Techniques: In Vivo, Derivative Assay, Immunohistochemistry

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: MiR-125b-5p targets HPRT1 in HNSCC. ( A ) qRT-PCR results analysis of HPRT1 hnRNA in HOK and HNSCC cells. ( B ) qRT-PCR and ( C ) Western blot analyses of HPRT1 in FaDu and SCC-4 cells transfected with miR-125b-5p mimic. ( D ) Pearson correlation analysis between miR-125b-5p and HPRT1 expression in HNSCC tissues. ( E ) Dual-luciferase reporting assay was used to detect the relative luciferase intensity of FaDu and SCC-4 cells co-transfected with miR-125b-5p mimic and wild-type/mutant HPRT1. ** P < 0.01, *** P < 0.001.

Article Snippet: Then, slides were incubated overnight at 4 °C with the rabbit anti-HPRT1 antibody (dilution:1:50, BBI, Shanghai, China).

Techniques: Quantitative RT-PCR, Western Blot, Transfection, Expressing, Luciferase, Mutagenesis

Journal: Heliyon

Article Title: Hypoxanthine guanine phosphoribosyltransferase 1, a target of miR-125b-5p, promotes cell proliferation and invasion in head and neck squamous cell carcinoma

doi: 10.1016/j.heliyon.2023.e20174

Figure Lengend Snippet: miR-125b-5p inhibits HNSCC cell proliferation and invasion by targeting HPRT1. ( A ) Western blot analysis of HPRT1 and MMP2 in HNSCC cells co-transfected with HPRT1 and miR-125b-5p. ( B ) CCK-8, ( C ) plate clone formation, and ( D ) transwell invasion assays were used to evaluate cell viability, proliferation, and invasion of HNSCC cells co-transfected with HPRT1 and miR-125b-5p. * P < 0.05, ** P < 0.01, *** P < 0.001. # P < 0.05, ## P < 0.01.

Article Snippet: Then, slides were incubated overnight at 4 °C with the rabbit anti-HPRT1 antibody (dilution:1:50, BBI, Shanghai, China).

Techniques: Western Blot, Transfection, CCK-8 Assay