Journal: BMC Pharmacology & Toxicology

Article Title: A novel nitidine chloride nanoparticle overcomes the stemness of CD133 + EPCAM + Huh7 hepatocellular carcinoma cells for liver cancer therapy

doi: 10.1186/s40360-022-00589-z

Figure Lengend Snippet: a . Representative organ images showing specific tumor targeting of rhodamine B isothiocyanate labeled TPGS-FA/NC nanoparticles 8 h post-injection into mice bearing Huh7 xenograft (T: tumor, Li: liver, H: heart, L: lung, K:kidney, S: spleen, and B:brain; Color scale: radiant efficiency, [p s − 1 cm − 2 sr − 1 ] [μWcm − 2 ] − 1 ). V b . Quantitative analysis of biodistribution in tumors and normal organs, quantified from the organ images. Intravenous treatment of nude mice bearing orthotopic Huh7 xenografts with TPGS-FA/NC nanoparticles (red) and control groups (turquoise: NC, fuchsia:5-Fu, blue: PBS) every other day for a total of five injections (4 mg kg − 1,NC per body weight, indicated by arrows). c . Mice body weight was monitored during the time course of treatments ( n = 5 biologically independent animals, statistics was calculated by two-tailed unpaired t-test presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, p = 4.3 × 10 − 3 ,3.4 × 10 − 3 and 5.0 × 10 − 4 comparing TPGS-FA/NC to NC,5-Fu and PBS, respectively). d . Representative images of liver cancer tumors harvested from mice after treatments * p < 0.05, ** p < 0.01, *** p < 0.001; p = 0.01, 8 × 10 − 4,and 2 × 10 − 4 comparing TPGS-FA/NC to NC, 5-Fu, and PBS, respectively. Source data are provided as a

Article Snippet: 5-fluorouracil (5-Fu) was purchased from MedChemExpress (MCE, Monmouth Junction, NJ, USA).

Techniques: Labeling, Injection, Control, Two Tailed Test

Journal: Oncotarget

Article Title: Identification of Mubritinib (TAK 165) as an inhibitor of KSHV driven primary effusion lymphoma via disruption of mitochondrial OXPHOS metabolism

doi: 10.18632/oncotarget.27815

Figure Lengend Snippet: ( A ) Graph of cell cycle kinetics data comparing Mubritinib (MUB) to cytarabine (CYT) and rapamycin (RAP). DMSO and camptothecin (CPT) were used as controls. PEL cells (BC1 and BCBL1) treated with 7.5 nM Mubritinib show a significant decrease in the total population of S and G2 cells that is not observed for BJAB or LCL352 cells. Neither cytarabine nor rapamycin showed similar selectivity for inhibition of PEL cell growth (S, G2) ( ** p < 0.001, * p < 0.05; Student’s T Test). ( B ) Cell cycle profiles comparing cells (BJAB, BC1, BCBL1, and LCL352) treated with DMSO and 15 nM Mubritinib measured by FACS flow cytometry analysis of propidium iodide staining. ( C ) Flow cytometry analysis of Annexin V/PI staining comparing cells (BJAB, BC1, BCBL1, LCL352) treated with DMSO and 15 nM Mubritinib. ( D ) Graphs summarizing the decrease in live cell populations observed from the Annexin V/PI experiment. Camptothecin (4 mM) and Cytarabin (1 mM) are shown for comparison. ( ** p < 0.001; Mann–Whitney). ( E ) ChIP-qPCR assay for LANA or IgG control in BCBL1 cells treated with Mubritinib (15 nM) for 72 hrs with primers for KSHV TR, control region a, or cellular Actin. ( F ) Western blot control of ChIP assays showing LANA, RTA, Actin, or gH2AX in BCBL1 cells at 48 h after addition of DMSO (–) or Mubritinib (15 nM) (+).

Article Snippet: Adrucil (228440010) and Cytarabine (449561000) were purchased from Acros Organics.

Techniques: Inhibition, Flow Cytometry, Staining, Comparison, MANN-WHITNEY, ChIP-qPCR, Control, Western Blot

Journal: Journal of Biological Chemistry

Article Title: The Caenorhabditis elegansmRNA 5′-Capping Enzyme

doi: 10.1074/jbc.m212101200

Figure Lengend Snippet: FIG. 5. The addition of the Ceg1-interacting region from Cet1 onto CEL-1 RTPase domain allows it to function in the presence of Ceg1. YSB719 was transformed with (left) pRSH-CEG1 or (right) pDB20H-MCE-(211–597). His isolates were subsequently transformed with the following LEU2/2 m plasmids: vector (pAD5); CET1 (pAD5- CET1); CET1-(1–225)-CEL-1-(13–221) (pAD5-CET1 (1–225)-CEL-1-(13– 221), expressing a fusion protein containing residues 1–225 of Cet1 and CEL-1-(13–221) tagged with the HA epitope); CET1-(1–265)-CEL-1-(13– 221) (pAD5-CET1 (1–265)-CEL-1-(13–221)); cet1–446-(1–265)-CEL-1-(13– 221) (pAD5-cet1–446-(1–265)-CEL-1-(13–221), expressing a fusion pro- tein consisting of residues 1–265 from the cet1–446 mutant (11) and CEL-1-(13–221) tagged with the HA epitope); and CEL-1-(13–221) (pAD5- CEL-1-(13–221)). Leu His transformants were grown in the presence of 5-FOA, and the plates were incubated for 3 days at 30 °C.

Article Snippet: Leu His transformants were grown in the presence of 5-FOA, and the plates were incubated for 3 days at 30 °C. at T E X A S U N IV ctober 16, 2014 http://w w w .jbc.org/ D ow nloaded from of the chimeric RTPases support viability of the same strain if MCE-(211–597) replaces Ceg1 as the GTase (Fig. 5, right), showing that these proteins are functional.

Techniques: Transformation Assay, Plasmid Preparation, Expressing, Mutagenesis, Incubation

Journal: Journal of Biological Chemistry

Article Title: The Caenorhabditis elegansmRNA 5′-Capping Enzyme

doi: 10.1074/jbc.m212101200

Figure Lengend Snippet: FIG. 6. RTPase activities of CEL-1-(13–221) mutants. A, purification of recombinant polyhistidine-tagged CEL-1-(13–221) protein. The peak fractions of the heparin-Sepharose CL-6B column were analyzed by SDS-PAGE and visualized by Coomassie Brilliant Blue staining (top panel) or immunoblotting with monoclonal anti-polyhistidine antibody (bottom panel). 400 and 50 ng of protein were loaded for the top and bottom panels, respectively. Lane 1, wild-type CEL-1-(13–221); lane 2, C136S; lane 3, R142K; lane 4, D76N; lane 5, E111Q; lane 6, D112N. For reasons that are not clear, the D76N mutant shows slightly altered mobility. B, RTPase assay. The wild-type and mutated CEL-1-(13–221) proteins were incubated at 30 °C for 10 min with 1 M termini of a [-32P]GTP-labeled 65 nucleotide RNA. Reaction mixtures were analyzed by thin layer chromatography (TLC) on polyethyleneimine-cellulose plates. Released phosphate was detected by autoradiography, and radioactive spots were cut out and quantitated by liquid scintillation counting. Relative amounts of released phosphate were plotted versus protein amount. C, in vivo analysis of CEL-1-(13–221) mutants by plasmid shuffling. YSB719 carrying pDB20H-MCE-(211–597) was transformed with the vector pAD5 or derivatives expressing the indicated CEL-1 alleles. Leu His transformants were tested for growth in the presence of 5-FOA. Plates were incubated for 4 days at 30 °C. D, immunoblot analysis of S. cerevisiae whole-cell extracts. YSB719 cells carrying pDB20H-MCE-(211–597) and CEL-1-(13–221) derivatives were grown in selective media but without shuffling out the CEG1/CET1 plasmid. Extracts were prepared, and 10-g protein was analyzed by SDS-PAGE and immunoblotting with the anti-HA antibody12CA5. Lane 1, pAD5; lane 2, pAD5-CEL-1-(13–221); lane 3, pAD5-CEL- 1-(13–221)C136S; lane 4, pAD5-CEL-1-(13–221)C136A; lane 5, pAD5-CEL-1-(13–221)R142K; lane 6, pAD5-CEL-1-(13–221)R142A; lane 7, pAD5- CEL-1-(13–221)D76N; lane 8, pAD5-CEL-1-(13–221)E111Q; lane 9, pAD5-CEL-1-(13–221)D112N.

Article Snippet: Leu His transformants were grown in the presence of 5-FOA, and the plates were incubated for 3 days at 30 °C. at T E X A S U N IV ctober 16, 2014 http://w w w .jbc.org/ D ow nloaded from of the chimeric RTPases support viability of the same strain if MCE-(211–597) replaces Ceg1 as the GTase (Fig. 5, right), showing that these proteins are functional.

Techniques: Purification, Recombinant, SDS Page, Staining, Western Blot, Mutagenesis, Incubation, Labeling, Thin Layer Chromatography, Autoradiography, In Vivo, Plasmid Preparation, Transformation Assay, Expressing