anti cmpk1  (Proteintech)

Journal: ACS pharmacology & translational science

Article Title: All Blood Brain Barrier Cell Types Demonstrate Capability to Influence Differential Tenofovir and Emtricitabine Metabolism and Transport in the Brain.

doi: 10.1021/acsptsci.4c00510

Figure Lengend Snippet: Figure 2. BBB cells differentially express TFV and FTC transporters and nucleotide-metabolizing kinases. Primary human BBB cell monoculture and hepatocyte lysates were processed for LC−MS/MS-based proteomics and protein concentrations for (A) ENT1, (B) MRP4, and (C) MRP1 TFV/FTC transporters, (D) AK2, (E) CKB, (F) PKLR, and (G) PKM TFV nucleotide-metabolizing kinases, as well as (H) CMPK1, (I) DCK, (J) PGK1, and (K) TK1 FTC nucleotide-metabolizing kinases, each with respective protein Western blots to confirm protein abundance in BBB cells. Concentrations of proteins of interest were measured by Proteomic Ruler. Four to five independent experiments with two LC−MS/MS injection replicates each were performed (represented as individual dots). Replicates with missing LC−MS/MS values were omitted from the plot. >2 independent experiments with missing values were reported as not reliably detected (ND) by proteomics analyses. Data represented as mean ± standard deviation. Statistical analysis was performed using Brown−Forsythe and Welch ANOVA or Kruskal−Wallis ANOVA by GraphPad software. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Western blots were performed as previously described.16 Blots were probed with antibodies with specificity to BCRP (NBP2-22124, Novus Bio, Centennial, CO), P-gp (PA5-61300, Invitrogen), MRP4 (12705, Cell Signaling Technology), MRP1 (Ab24102, Abcam), ENT1 (PA5-116451, Invitrogen), PGK1 (PA528612, Invitrogen), AK2 (11014-1-AP, Proteintech, Rosemont, IL), CKB/CKM (15137-1-AP, Proteintech), TK (15691-1-AP, Proteintech), CMPK (11360-1-AP, Proteintech), DCK (17758-1-AP, Proteintech), PKM1 (15821-1-AP, Proteintech), and PKLR (2456-1-AP, Proteintech), overnight at 4 °C, washed with TBS-T, and probed with the appropriate secondary antibody (ab97023 and Ab97051, Abcam) for 1 h at room temperature.

Techniques: Liquid Chromatography with Mass Spectroscopy, Western Blot, Quantitative Proteomics, Injection, Standard Deviation, Software

Journal: ACS pharmacology & translational science

Article Title: All Blood Brain Barrier Cell Types Demonstrate Capability to Influence Differential Tenofovir and Emtricitabine Metabolism and Transport in the Brain.

doi: 10.1021/acsptsci.4c00510

Figure Lengend Snippet: Figure 4. BBB TFV/FTC nucleotide-metabolizing kinases and transporters are impacted in a cell-dependent manner by ART and HIV. Primary human pericyte and astrocyte monocultures were exposed to ART (10 μM TFV, 10 μM FTC, and 10 μM DTG), HIV (5 ng/mL), or HIV + ART (10 μM TFV, 10 μM FTC, 10 μM DTG, and 5 ng/mL HIV) for 24 h at 37 °C, 5% CO2. Treatment with a vehicle was used as a control. Pericytes and astrocytes were lysed and processed for proteomics analyses. Concentrations of proteins of interest were measured by Proteomic Ruler. After exposure, changes in protein concentration were assessed in (A−G) astrocytes (astro) for (A) AK2, (B) CMPK1, (C) ENT1, (D) P-gp, (E) PGK1, (F) MRP4, and (G) MRP1. Similarly, changes in protein concentration were assessed in (H−L) pericytes (per) for (H) CKB, (I) MRP4, (J) MRP1, (K) PGK1, and (L) TK1. The significant changes (A−L) in the concentrations of TFV and FTC transporters and metabolizing enzymes after ART, HIV, or HIV + ART exposure relative to vehicle were (M) summarized for astrocytes (astro) and pericytes (per). An arrow next to each protein denotes an increase (upward arrow) or decrease (downward arrow) in protein concentration after ART, HIV, or HIV + ART exposure relative to the vehicle. Four independent experiments with two LC−MS/MS injection replicates each were performed per condition (represented as individual dots). Replicates with missing LC−MS/MS values were omitted from the plot. >2 independent experiments with missing values were reported as not reliably detected (ND) by proteomics analyses. Data represented as mean ± standard deviation. Statistical analysis was performed using Brown−Forsythe and Welch ANOVA or Kruskal−Wallis ANOVA by GraphPad software. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

Article Snippet: Western blots were performed as previously described.16 Blots were probed with antibodies with specificity to BCRP (NBP2-22124, Novus Bio, Centennial, CO), P-gp (PA5-61300, Invitrogen), MRP4 (12705, Cell Signaling Technology), MRP1 (Ab24102, Abcam), ENT1 (PA5-116451, Invitrogen), PGK1 (PA528612, Invitrogen), AK2 (11014-1-AP, Proteintech, Rosemont, IL), CKB/CKM (15137-1-AP, Proteintech), TK (15691-1-AP, Proteintech), CMPK (11360-1-AP, Proteintech), DCK (17758-1-AP, Proteintech), PKM1 (15821-1-AP, Proteintech), and PKLR (2456-1-AP, Proteintech), overnight at 4 °C, washed with TBS-T, and probed with the appropriate secondary antibody (ab97023 and Ab97051, Abcam) for 1 h at room temperature.

Techniques: Control, Protein Concentration, Liquid Chromatography with Mass Spectroscopy, Injection, Standard Deviation, Software

Journal: iScience

Article Title: Exosomal miRNA 16-5p/29a-3p from pancreatic cancer induce adipose atrophy by inhibiting adipogenesis and promoting lipolysis

doi: 10.1016/j.isci.2024.110346

Figure Lengend Snippet:

Article Snippet: Anti-CD81 (sc-166029) antibody was purchased from Santa Cruz, Dallas, TX, USA; anti-C/EBPβ (cat:606202) was obtained from BioLegend, San Diego, CA, USA; anti-TSG101 (GTX70255), anti-Erlin2 (GTX106277), anti-CMPK1 (GTX105566), anti-MCT1 (GTX-631643), anti-Cse1L (GTX103005), anti-RFP (GTX127897) and anti-GAPDH (GTX100118) antibodies were from GeneTex, Irvine, CA, USA; anti-Rab27a (Cat:17817-1-AP) and anti-Rab27b (Cat:13412-1-AP) antibodies were obtained from Proteintech, USA; anti-ATGL (2138s) and anti-PPARγ (2443s) antibodies were gained from Cell Signaling Technology, Danvers, MA, USA.

Techniques: Recombinant, Lysis, Transfection, Fluorescence, SYBR Green Assay, Isolation, Luciferase, Mass Spectrometry, Negative Control, Software

Journal: Biology

Article Title: The Metabolic Activation of Sofosbuvir Is Impaired in an Experimental Model of NAFLD

doi: 10.3390/biology11050693

Figure Lengend Snippet: Sequences of the Primers Used in the Study for qRT-PCR Experiments.

Article Snippet: After a blocking step performed with 10% skim milk in TBS-T, the membrane was incubated overnight at 4 degrees with a primary anti-UMP-CMPK1 antibody (Santa Cruz Biotechnology; Dallas, TX, USA, diluted 1:500), and then with a secondary anti-mouse HRP-conjugated antibody (LGC Seracare; Milford, MA, USA, diluted 1:100).

Techniques:

Journal: Biology

Article Title: The Metabolic Activation of Sofosbuvir Is Impaired in an Experimental Model of NAFLD

doi: 10.3390/biology11050693

Figure Lengend Snippet: mRNA expression of the enzymes UMP-CMPK1 ( A ) and NDPK ( B ), responsible for the activation of sofosbuvir to the active compound GS-331007-TP. Protein expression of the enzyme UMP-CMPK1 ( C ). A representative Western Blot is shown in ( D ). Results are reported as means and S.E.M. of 6 animals per group. * p < 0.05 vs. healthy rats.

Article Snippet: After a blocking step performed with 10% skim milk in TBS-T, the membrane was incubated overnight at 4 degrees with a primary anti-UMP-CMPK1 antibody (Santa Cruz Biotechnology; Dallas, TX, USA, diluted 1:500), and then with a secondary anti-mouse HRP-conjugated antibody (LGC Seracare; Milford, MA, USA, diluted 1:100).

Techniques: Expressing, Activation Assay, Western Blot