Journal: Molecular Cancer Research

Article Title: 5-Hydroxy-2-Methyl-1,4-Naphthoquinone, a Vitamin K3 Analogue, Suppresses STAT3 Activation Pathway through Induction of Protein Tyrosine Phosphatase, SHP-1: Potential Role in Chemosensitization

doi: 10.1158/1541-7786.mcr-09-0257

Figure Lengend Snippet: FIGURE 6. A. Overexpression of constitutive STAT3 rescues A293 cells from plumbagin-induced cytotoxicity. First, A293 cells were transfected with constitutive STAT3 plasmid. After 24 h of transfection, the cells were treated with 2.5 μmol/L plumbagin for 24 h, the cytotoxicity was determined by Live/Dead assay, and 20 random fields were counted. B. Plumbagin potentiates the apoptotic effect of thalidomide and bortezomib. U266 cells (1 × 106) were treated with 2.5 μmol/L plumbagin, and 10 μg/mL thalidomide or 20 nmol/L bortezomib alone or in combination for 24 h at 37°C. Cells were stained with a Live/Dead assay reagent for 30 min and then analyzed under a fluorescence microscope. The results shown are representative of two independent experiments.

Article Snippet: Bortezomib (Velcade, PS-341) was obtained from Millennium, and thalidomide was from Tocris Cookson.

Techniques: Over Expression, Transfection, Plasmid Preparation, Live Dead Assay, Staining, Fluorescence, Microscopy

Journal: bioRxiv

Article Title: Magnetoelectric Nanodiscs Enable Wireless Transgene-Free Neuromodulation

doi: 10.1101/2023.12.24.573272

Figure Lengend Snippet: Relative GCaMP6s fluorescence change (ΔF/F 0 ) in hippocampal neurons decorated with MENDs before (left) and after (right) magnetic field application. Scale bars: 150 µm. b, Individual (top) and average (bottom) of the relative GCaMP6s fluorescence change (ΔF/F 0 ) of the hippocampal neurons in response to 10 mT AMF with 1 kHz frequency and 220 mT OFM. c, Individual and d, average traces of GCaMP6s ΔF/F 0 in 300 hippocampal neurons decorated with MENDs in response to 10 mT AMF with frequencies 100, 150, 250, 500, and 1000 Hz (OMF magnitude 220 mT). The grey and magenta dashed lines indicate the beginning and end of MF stimulation, respectively. e, Individual cell (top) and mean (bottom) GCaMP6s fluorescence changes in 300 neurons in response to 2 s MF epochs applied at varying intervals (OMF 220 mT; AMF 150 Hz, 10 mT). f, Number of GCaMP6s fluorescence peaks as a function of stimulation epoch length for rest intervals of 10, 30, 60, 90, and 120 s (OMF 220 mT; AMF 150 Hz, 10 mT). g, Individual cell (top) and mean (bottom) GCaMP6s fluorescence changes in response to 2 s MF epochs at 40 s intervals for 0.75 µg mm −2 MEND density. h, i, Fluorescent image of a Live-Dead assay in neurons before and after 3 cycles of MF at h, 1 µg mm −2 and i, 0.75 µg mm −2 MEND densities. j, The change in live cell ratio (normalized to the total number of the cells counted with Hoest staining) following 3 cycles of MF for neurons decorated with different MEND densities. Statistical significance was tested via one-way ANOVA and Tukey’s multiple comparison tests (n = 5 plates per condition, P=3.79×10 −7 for 1 µg mm −2 ; P=0.79 for 0.794 µg mm −2 ; P=0.998 for 0.75 µg mm −2 ; ****P≤0.0001, n.s. P>0.05).

Article Snippet: 4 days following seeding, the neurons were transduced with 1 μL of an adeno-associated virus serotype 9 (AAV9) carrying a fluorescent calcium ion indicator GCaMP6s under a pan-neuronal human synapsin (hSyn) promoter (AAV9-hSyn::GCaMP6s, Addgene viral prep #100843-AAV9, >1×10 13 IU/μL).

Techniques: Fluorescence, Live Dead Assay, Staining, Comparison

Journal: bioRxiv

Article Title: Magnetoelectric Nanodiscs Enable Wireless Transgene-Free Neuromodulation

doi: 10.1101/2023.12.24.573272

Figure Lengend Snippet: Illustration of stimulation mechanism combining cable model and repeated excitation of neural activity, where d is spacing between MEND particles, a is cell radius, Δ V is the change in membrane potential per half-period of an AMF, and V 0 is the voltage generated by a single MEND. b, Calculated membrane potential change Δ V for every half-period of an AMF, as a function of V 0 and d . c, Simulated membrane potential V ( t ) after AMF onset to activate MENDs with V 0 = 0.03 mV for varying a values. The threshold potential for action potential firing, –55 mV, is indicated with the dashed line. d, Simulated membrane potential at a time t = 2 s after onset of magnetic field as a function of d , for varying V 0 values. V 0 =0.03 mV is the measured value for MEND particles in this study, highlighted in red. e, Time to reach threshold membrane potential (–55 mV) from the resting potential (–75 mV) as a function of AMF frequency ƒ AMF for varying V 0 values. f, ( i-iii ) SEM images showing MENDs decorating cultured hippocampal neurons. (ii) A higher magnification image of the area marked by a box in panel (i). (iii) MENDs on the neuron surface shaded in blue as identified by the presence of iron, titanium, and barium in energy dispersive X-ray spectroscopy. Scale bars: 20 µm (i), 5 µm (ii), 100 nm (iii). g, h, GCaMP6s fluorescence changes in neurons decorated with MEND following 2 s stimulations (OMF 220 mT; AMF 150 Hz, 10 mT, marked by vertical grey bars) in the presence of g, tetrodotoxin (TTX, 1 µM, green) or h, a cocktail of (2R)-amino-5-phosphonovaleric acid (AP5, 100 µM) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 20 µM) (blue).

Article Snippet: 4 days following seeding, the neurons were transduced with 1 μL of an adeno-associated virus serotype 9 (AAV9) carrying a fluorescent calcium ion indicator GCaMP6s under a pan-neuronal human synapsin (hSyn) promoter (AAV9-hSyn::GCaMP6s, Addgene viral prep #100843-AAV9, >1×10 13 IU/μL).

Techniques: Activity Assay, Membrane, Generated, Cell Culture, Spectroscopy, Fluorescence

Journal: bioRxiv

Article Title: Magnetoelectric Nanodiscs Enable Wireless Transgene-Free Neuromodulation

doi: 10.1101/2023.12.24.573272

Figure Lengend Snippet: Schematic of MEND-mediated neuromodulation. MEND particles were injected into the mouse ventral tegmental area (VTA), and the mice were placed inside a permanent magnet field providing OMF of 220 mT and a surrounding solenoid providing AMF with an amplitude 10 mT and a frequency 150 Hz. b, Confocal images of c-Fos-expressing neurons among DAPI-marked cells in the VTA. Top left: MENDs (1.5 mg/ml) with (+) magnetic stimulation (ON); top right: MEND particles (1.5 mg/ml) without (–) magnetic stimulation (OFF); bottom left: control MNDs (1.5 mg/ml) + magnetic stimulation; bottom right: MENDs (0.5 mg/ml) + magnetic stimulation. Scale bars are 25 µm. c, Quantification of c-Fos positive neurons for the conditions shown in (B) as well as the subjects injected with PBS and exposed to magnetic field. d, Confocal images and e, quantification of c-Fos positive neurons in the medial prefrontal cortex (mPFC) for the same conditions shown in (B) and (C). f, Confocal images and g, quantification of c-Fos positive neurons in the nucleus accumbens (NAc) for the same conditions shown in b and c . d, f, Scale bars are 100 µm. c, e, g, Statistical significance was tested via one-way ANOVA and Tukey’s multiple comparison tests (n = 6 mice per condition, VTA F 3,20 =93.2 P=1.21×10 −12 ; mPFC F 3,20 =60.81 P=1.21×10 −11 ; VTA F 3,20 =62.03 P=1.29×10 −12 ; ****P≤0.0001). h, Representative confocal images showing expression of GCaMP6s in the VTA. Scale bar: 50 µm. i Fiber photometry recordings of relative GCaMP6s fluorescence change (ΔF/F 0 ) in the VTA of anaesthetized mice injected with MENDs in the same region. (Top) Individual trial ΔF/F 0 . (Bottom) Average ΔF/F 0 across trials shown above. Solid line represents mean and shaded areas mark s.e.m. (n=4 mice, 10-15 trials per mouse). The grey square box indicates magnetic field epochs (2s, OMF 220 mT, AMF 10 mT, 150 Hz). j Schematic of the place preference behavioral apparatus (top) and experimental timeline (bottom). k, Time spent in the stimulation chamber out of a total test time of 600 s, for pre-(Day 1, open markers) and post-(Day 5, solid markers) learning. Paired t-test was performed for MEND (n=11) and MND (n=7) groups, and Wilcoxon signed-rank test was performed for PBS (n=7) group because the data did not follow normal distribution. ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05, n.s. P>0.05). l, The change in time spent in stimulation chamber between Day 1 and Day 5. (One-way ANOVA with Tukey’s post-hoc comparison test, ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05, n.s. P>0.05.)

Article Snippet: 4 days following seeding, the neurons were transduced with 1 μL of an adeno-associated virus serotype 9 (AAV9) carrying a fluorescent calcium ion indicator GCaMP6s under a pan-neuronal human synapsin (hSyn) promoter (AAV9-hSyn::GCaMP6s, Addgene viral prep #100843-AAV9, >1×10 13 IU/μL).

Techniques: Injection, Expressing, Control, Comparison, Fluorescence