rabbit anti agmat novus biological  (Novus Biologicals)

Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet:

Article Snippet: Rabbit anti-AGMAT , Novus Biological , Cat#1-82080.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Luciferase, Reporter Assay, RNA Sequencing, Control, Mutagenesis, CRISPR, Plasmid Preparation, shRNA, Software

Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet: ARG1, AGMAT, arginine, and RBM39 in human HCC patients (A) Schematic representation of arginine and polyamine metabolism in HCC patients. Boxes below enzymes indicate changes in mRNA (left box) and protein (right box) levels in human HCC tumors (T) compared to paired non-tumor (NT) biopsies, respectively. Color coding according to level of log 2 fold-change as indicated. “?” indicates unknown identity. Tumor aggressiveness is indicated by Edmondson-Steiner grade low (Edm. low, grade I and II) and high (Edm. high, grade III and IV). n = 73 (Edm. low) and n = 49 (Edm. high) for mRNA; n = 30 (Edm. low) and n = 21 (Edm. high) for protein. (B) Immunoblots of ARG1, AGMAT, RBM39, and ASNS in paired non-tumor (NT) and tumor (T) tissues of five HCC patients. Calnexin serves as loading control. (C) Tissue microarray for ARG1 and AGMAT. ARG1, normal liver n = 58, HCC n = 160; AGMAT, normal liver n = 49, HCC n = 142. (D) Representative IHC of ARG1 and AGMAT of an HCC patient (from C). Non-tumor, NT; tumor, T. (E) Kaplan-Meier survival estimate curve for The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA-LIHC) patients ranked by expression of ARG1 and AGMAT . n = 89 (low), n = 109 (normal). (F) Urea cycle metabolites in tumors (T) relative to paired non-tumor (NT) liver tissues (log 2 ratio). n = 11. (G) Immunoblots of RBM39 in tumor lysate (Input) and elution after purification with leucine (Leu)- or arginine (Arg)-coupled agarose beads from three HCC patients. Calnexin serves as input and negative control. (H) Dose-response curve of 20 HCC patient-derived organoids treated with indisulam. Data are presented as the percentage of control DMSO-treated tumor organoids. (I) Model. In liver cancer cells, loss of ARG1 and AGMAT preserves arginine, which in turn binds RBM39 to promote metabolic reprogramming. Arginine-RBM39-mediated ASNS expression further enhances arginine uptake. Trsx, transcription. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001 by unpaired t test (C), log rank test (E), and multiple t test (F).

Article Snippet: Rabbit anti-AGMAT (for IHC) , Sigma , Cat#PA5-55311.

Techniques: Western Blot, Microarray, Expressing, Purification, Negative Control, Derivative Assay

Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet: ARG1 and AGMAT are decreased and arginine, RBM39, and ASNS are increased in HCC patient tumors that are sensitive to RBM39 depletion by indisulam, related to Figure 7 (A) RBM39 mRNA levels in liver tumor tissue (T) from HCC patients compared to adjacent non-tumor tissue (NT), displayed as log 2 ratio. n = 73 (Edm. low), n = 49 (Edm. high). (B) RBM39 protein levels in liver tumor tissue (T) from HCC patients compared to adjacent non-tumor tissue (NT), displayed as log 2 ratio. n = 30 (Edm. low), n = 21 (Edm. high). (C) ASNS mRNA levels in liver tumor tissue (T) from HCC patients compared to adjacent non-tumor tissue (NT), displayed as log 2 ratio. n = 73 (Edm. low), n = 49 (Edm. high). (D) ASNS protein levels in liver tumor tissue (T) from HCC patients compared to adjacent non-tumor tissue (NT), displayed as log 2 ratio, if applicable. BW, black-and-white, i.e., only detected in tumor tissues. n = 3 (Edm. low), n = 8 (Edm. high). (E) Staging of ARG1 and AGMAT IHC staining in tissue micro array. (F) mRNA expression of ARG1 , AGMAT , RBM39 , and ASNS in early-stage HCC (data from Jiang et al. ). log 2 fold-change tumor (T) relative to non-tumor (NT) tissues. n = 35. (G) Kaplan-Meier survival estimate curve for TCGA-LIHC patients ranked by expression of ARG1 . n = 135 (low), n =155 (normal). (H) Kaplan-Meier survival estimate curve for TCGA-LIHC patients ranked by expression of AGMAT . n = 136 (low), n = 158 (normal). (I) Polyamine species in tumors (T) relative to paired non-tumor (NT) liver tissues (log 2 ratio). n = 11. (J) Arginine content in paired non-tumor (NT) and tumor (T) tissues of HCC patients. n = 10. (K) Total polyamine content in paired non-tumor (NT) and tumor (T) tissues of HCC patients. n = 10. (L) Volcano plot of the −log 10 (adjusted p value) against the log 2 fold-change of 600 proteins identified by MS (in minimum 2 out of 3 samples) after purification from HCC tissues by arginine (Arg)- compared to leucine (Leu)-coupled agarose beads. Red dot highlights RBM39. (M) Dose-response curve of 20 HCC patient-derived organoids treated with sorafenib. Data are presented as the percentage of control DMSO-treated tumor organoids. (N) IC 50 of indisulam- and sorafenib-treated HCC patient-derived organoids. n = 20. (O and P) Rbm39 and Asns mRNA levels in embryonic day 14 (E14), E18, and adult mouse liver as reads per kilobase of exon per million reads mapped (RPKM). Data from NBCI Gene. n.s. = not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 by paired t test (A–C, J, K, and N), multiple t test (F and I), and log rank test (G and H).

Article Snippet: Rabbit anti-AGMAT (for IHC) , Sigma , Cat#PA5-55311.

Techniques: Immunohistochemistry, Microarray, Expressing, Purification, Derivative Assay

Journal: Cell

Article Title: Arginine reprograms metabolism in liver cancer via RBM39

doi: 10.1016/j.cell.2023.09.011

Figure Lengend Snippet:

Article Snippet: Rabbit anti-AGMAT (for IHC) , Sigma , Cat#PA5-55311.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Luciferase, Reporter Assay, RNA Sequencing Assay, Mutagenesis, CRISPR, Plasmid Preparation, shRNA, Software

Journal: Journal of Animal Science and Biotechnology

Article Title: Pre-implantation exogenous progesterone and pregnancy in sheep: I. polyamines, nutrient transport, and progestamedins

doi: 10.1186/s40104-021-00554-6

Figure Lengend Snippet: Endometrial expression of mRNAs for progestamedins, enzymes involved in polyamine synthesis, and nutrient transporters. Expression of FGF10 ( d ), AZIN2 (H), SLC6A9 ( q ), and SLC1A4 ( r ) were affected by treatment. Expression of SLC2A1 ( n ), SLC5A1 ( o ), and SLC7A1 ( p ) were affected by day. Expression of FGF10 ( b ), ODC1 ( e ), SLC2A1 ( i ), SLC5A1 ( j ), SLC7A1 ( k ), and SLC6A9 ( l ) were affected by a day × treatment interaction. Expression of FGF7 ( a ), HGF ( c ), AZIN2 ( f ) , and AGMAT ( g ) were not affected by a day × treatment interaction. There was a tendency for the endometrial expression of SLC1A4 ( m ) to be affected by a day × treatment interaction. Data are presented as LSM ± SEM. Different means are indicated by different letters. Only endometria from ewes which were considered pregnant with normally developed conceptuse were utilized for these analyses

Article Snippet: AGMAT protein was detected using a primary rabbit polyclonal antibody to AGMAT (Abcam 231,894; Cambridge, UK) at a final dilution of 1:250.

Techniques: Expressing

Journal: Journal of Animal Science and Biotechnology

Article Title: Pre-implantation exogenous progesterone and pregnancy in sheep: I. polyamines, nutrient transport, and progestamedins

doi: 10.1186/s40104-021-00554-6

Figure Lengend Snippet: Localization of endometrial AGMAT. AGMAT proteins localized to uterine luminal epithelia (LE), superficial glandular epithelia (sGE), GE, and stromal cells in both P4- and CO-treated ewes on day 9 and 12 of pregnancy ( a ). Intensity of AGMAT staining in the GE was decreased in endometria at day 12 compared to day 9 ( P < 0.01) ( b ). A day × treatment interaction for the intensity of AGMAT staining in the LE was observed ( P < 0.001), with increased intensity of AGMAT staining observed in the LE from P4-treated ewes compared to CO-treated ewes at day 12. Scale bar represents 100 μm. Only endometria from ewes which were considered pregnant with normally developed conceptuses were utilized for these analyses

Article Snippet: AGMAT protein was detected using a primary rabbit polyclonal antibody to AGMAT (Abcam 231,894; Cambridge, UK) at a final dilution of 1:250.

Techniques: Staining