cell culture human monocytic u937 cells  (ATCC)

Journal: Journal of proteome research

Article Title: Simple and Efficient Microsolid-Phase Extraction Tip-Based Sample Preparation Workflow to Enable Sensitive Proteomic Profiling of Limited Samples (200 to 10,000 Cells)

doi: 10.1021/acs.jproteome.0c00890

Figure Lengend Snippet: Comparison of Lys-C and trypsin addition sequences by OmSET sample preparation for processing 2000 U937 cells. Numbers of protein groups (A) and peptide groups (B) identified from aliquots equivalent to ~200 injected cells for three tested sequences of enzyme additions: addition of Lys-C (estimated enzyme to substrate ratio of 1:10) and trypsin (1:10) simultaneously (“simult”), addition of Lys-C (1:10) and trypsin (1:10) simultaneously followed by a readdition of trypsin (1:20) after 1 h (“trypsin readd”), and pretreatment with Lys-C (1:10) for 30 min before the addition of trypsin (1:10) (“Lys-C preadd”). Error bars indicate standard deviation from sample processing triplicates with LC–MS technical duplicates each (N = 6 in total). Percentages of 0 (yellow), 1 (light blue), and 2 miscleavages (gray) for peptide sequence matches are shown in panel (C). The max number of shown miscleavages was 2.

Article Snippet: U937 Cell Culture Human monocyte U937 cells (ATCC, Manassas, VA) were grown in the RPMI 1640 medium with l -glutamine supplemented with 10% FBS and 1% P/S at 37 °C and 5% CO 2 .

Techniques: Comparison, Sample Prep, Injection, Standard Deviation, Liquid Chromatography with Mass Spectroscopy, Sequencing

Journal: Journal of proteome research

Article Title: Simple and Efficient Microsolid-Phase Extraction Tip-Based Sample Preparation Workflow to Enable Sensitive Proteomic Profiling of Limited Samples (200 to 10,000 Cells)

doi: 10.1021/acs.jproteome.0c00890

Figure Lengend Snippet: Evaluation of depth of proteomic profiling for 10 to 500 U937 cells by OmSET sample preparation. Numbers of protein groups (A) and peptide groups (B) for 0, 10, 100, and 500 cell equivalents injected from 0, 200, 2000, and 10,000 cells processed, respectively. An aliquot of PBS was processed in parallel and used as a negative control (0 cell). Solid bars represent identifications using database searching only, while outlined bars show identifications using database and spectral library searching in combination. Error bars show standard deviation from triplicate LC–MS runs of sample processing duplicates (N = 6 in total). Numbers of protein groups (C) and peptide groups (D) identified from equivalent samples prepared using a conventional in-solution approach. Venn diagram (E) shows the overlap in protein group identifications between the three cell loading amounts and the blank sample processed using OmSET.

Article Snippet: U937 Cell Culture Human monocyte U937 cells (ATCC, Manassas, VA) were grown in the RPMI 1640 medium with l -glutamine supplemented with 10% FBS and 1% P/S at 37 °C and 5% CO 2 .

Techniques: Sample Prep, Injection, Negative Control, Standard Deviation, Liquid Chromatography with Mass Spectroscopy

Journal: Journal of proteome research

Article Title: Simple and Efficient Microsolid-Phase Extraction Tip-Based Sample Preparation Workflow to Enable Sensitive Proteomic Profiling of Limited Samples (200 to 10,000 Cells)

doi: 10.1021/acs.jproteome.0c00890

Figure Lengend Snippet: Comparison of label-free quantitation results from ~10, ~100, and ~500 U937 cells. Pairwise correlations of protein abundances for LC–MS runs of the same sample for aliquots equivalent to ~10 cells (A), ~100 cells (B), and ~500 cells (C). Pairwise correlations of protein abundances for LC–MS runs of different sample processing replicates for aliquots equivalent to ~10 cells (D), ~100 cells (E), and ~500 cells (F). Examples of low (G), medium (H), and high (I) abundance proteins from the comparison of protein abundances to the number of cells analyzed. Gene names are given for each protein example. Distribution of R2 values from the bottom 25%, top 25%, and all abundances for protein abundances compared to the number of cells analyzed (J). R2 values are given as mean ± SD.

Article Snippet: U937 Cell Culture Human monocyte U937 cells (ATCC, Manassas, VA) were grown in the RPMI 1640 medium with l -glutamine supplemented with 10% FBS and 1% P/S at 37 °C and 5% CO 2 .

Techniques: Comparison, Quantitation Assay, Liquid Chromatography with Mass Spectroscopy

Journal: British Journal of Pharmacology

Article Title: The antinociceptive triterpene β-amyrin inhibits 2-arachidonoylglycerol (2-AG) hydrolysis without directly targeting cannabinoid receptors

doi: 10.1111/j.1476-5381.2012.02059.x

Figure Lengend Snippet: The effect of amyrins on 2-AG trafficking and breakdown in intact U937 cells. (A) [3H]-2-AG intracellular levels and (B) [3H]-glycerol formation in U937 cells pretreated with amyrins, JZL184 or vehicle and then incubated with 1 µM 2-AG/[3H]2-AG mix for 5 min. The effect of pristimerin and MAFP is shown only at a concentration of 10 µM. The radioactive counts in the vehicle controls were approximately 9000 and 300 dpm for [3H]-glycerol formation and intracellular [3H]-2-AG, respectively. Data show means ± SEM, n= 9 from three independent experiments. *P < 0.05, **P < 0.01, significantly different from vehicle-treated samples (=100%).

Article Snippet: Cell culture Human leukemia monocytic lymphoma U937 cells were purchased from American Type Culture Collection (Manassas, VA, USA) and were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum, 1 g·mL −1 fungizone (amphotericin B), 100 units·mL −1 penicillin, 100 g·mL −1 streptomycin and 2 mM L-glutamine (all from Invitrogen, Carlsbad, CA, USA).

Techniques: Incubation, Concentration Assay

Journal: British Journal of Pharmacology

Article Title: The antinociceptive triterpene β-amyrin inhibits 2-arachidonoylglycerol (2-AG) hydrolysis without directly targeting cannabinoid receptors

doi: 10.1111/j.1476-5381.2012.02059.x

Figure Lengend Snippet: The effect of amyrins on 2-AG uptake in intact U937 cells pretreated with 10 µM JZL184. Intracellular [3H]-2-AG levels in U937 cells pretreated with (A) β-amyrin plus JZL184 or β-amyrin plus vehicle, (B) α-amyrin plus JZL184 or α-amyrin plus vehicle incubated with 1 µM 2-AG/[3H]-2-AG for 5 min. Data show mean values ± SEM, n= 12 from four independent experiments. *P < 0.05, **P < 0.01, significantly different from vehicle control (=100%). #P < 0.05, ##P < 0.01, JZL184 plus amyrin-treated cells significantly different from amyrin-treated cells (=100%).

Article Snippet: Cell culture Human leukemia monocytic lymphoma U937 cells were purchased from American Type Culture Collection (Manassas, VA, USA) and were grown in RPMI 1640 medium supplemented with 10% fetal bovine serum, 1 g·mL −1 fungizone (amphotericin B), 100 units·mL −1 penicillin, 100 g·mL −1 streptomycin and 2 mM L-glutamine (all from Invitrogen, Carlsbad, CA, USA).

Techniques: Incubation, Control