Journal: bioRxiv

Article Title: The polyamine naphthyl-acetyl spermine trihydrochloride (NASPM) lacks specificity for Ca 2+ -permeable AMPA receptors and suppresses seizure like activity in human brain tissue by inhibition of NMDA receptors

doi: 10.1101/2025.05.14.653650

Figure Lengend Snippet: A. Currents recorded from ND7/23 cells expressing GluN1/GluN2A receptors, at –60 mV, –30 mV and +30 mV, respectively, upon application of NMDA (100 µM, black traces) or NMDA + NASPM (8.7 μM, 26.1 μM, 87 μM, color gradient traces) in a Mg 2+ -free condition. B. Normalized GluN1/GluN2A receptor peak currents, categorized by membrane potential and concentration, with upper and lower quartiles, mean and standard deviation. C. Estimated concentration dependencies of GluN1/GluN2A block by NASPM. Average normalized GluN1/GluN2A peak currents were fitted with a rectangular hyperbolic function, assuming direct occlusion of the ion channel pore by a single NASPM molecule. The IC 50 values from the fits were 3.29 ± 0.54 µM, 23.6 ± 1.1 µM, and 56.1 ± 7.4 µM at –60 mV, –30 mV, and +30 mV.

Article Snippet: They were passaged twice a week and seeded onto glass coverslips in 60mm Petri dishes two days prior to experiments. pCI-EGFP-NR2a wt (Addgene plasmid #45445, http://n2t.net/addgene:45445 ; RRID:Addgene_45445) and pCI-EGFP-NR1 wt (Addgene plasmid #45446, http://n2t.net/addgene:45446 ; RRID:Addgene_45446) were gifts from Andres Barria & Robert Malinow (Barria et Malinow, 2002).

Techniques: Expressing, Membrane, Concentration Assay, Standard Deviation, Blocking Assay

Journal: Pharmaceutics

Article Title: Novel Dimer Derivatives of PF-543 as Potential Antitumor Agents for the Treatment of Non-Small Cell Lung Cancer.

doi: 10.3390/pharmaceutics14102035

Figure Lengend Snippet: Figure 3. Effects of PF-543 and PF-543 derivatives on A549 cells biological result; (a) Activities of SK1 and SK2 at 10 µM concentrations of PF-543 and PF-543 derivatives; (b) Protein was extracted and detected SK1, SK2, and β-actin protein levels by Western blotting. Each protein level was normalized by β-actin. Western blot was performed in three independent experiments; (c) The mRNA levels of SK1 and SK2 were quantified by qRT-PCR by treating A549 cells with PF-543 or PF-543 derivatives at 5 and 10 µM for 24 h. SK1 and SK2 mRNA was normalized to cyclophilin; (d) Sphingosine, ceramide, and S1P levels were measured in A549 cells with 5 and 10 µM PF-543 and PF-543 derivatives. The data are expressed as the mean ± S.D. * p < 0.05, *** p < 0.001 compared with control group.

Article Snippet: Control small interfering RNA (siControl) was obtained from Bioneer (Daejeon, Korea), while SK1 (sc-44114) and SK2 (sc-39225) siRNA (siSK1 and siSK2) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Western Blot, Quantitative RT-PCR, Control

Journal: Pharmaceutics

Article Title: Novel Dimer Derivatives of PF-543 as Potential Antitumor Agents for the Treatment of Non-Small Cell Lung Cancer.

doi: 10.3390/pharmaceutics14102035

Figure Lengend Snippet: Figure 5. Cell cytotoxic effect of Compounds 2 and 4 by inhibiting SK1 and SK2 expression. A549 cells were transiently transfected with control siRNA or SK1, SK2 siRNA for 24 h; (a) The protein expression level of SK1 and SK2 was confirmed by Western blot; (b–d) Cells transfected with siSK1, siSK2 or siSK1 + siSK2 were treated with Compounds 2 and 4 and FP-543 for 24 h, respectively, and cell viability was measured using EZ-CYTOX. Data are expressed as the mean ± S.D. *** p < 0.001 compared with control group.

Article Snippet: Control small interfering RNA (siControl) was obtained from Bioneer (Daejeon, Korea), while SK1 (sc-44114) and SK2 (sc-39225) siRNA (siSK1 and siSK2) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Expressing, Transfection, Control, Western Blot