Journal: Research

Article Title: Targeting the LSD1-G9a-ER Stress Pathway as a Novel Therapeutic Strategy for Esophageal Squamous Cell Carcinoma

doi: 10.34133/2022/9814652

Figure Lengend Snippet: Integrative transcriptomics analyses of ESCC cells following pharmacological or genetic inhibition of LSD1 and G9a. (a) RNA-seq analysis was performed on ESCC cells in which either LSD1 expression was knocked down using shRNA or LSD1 was inhibited with SP2509; differentially expressed genes were determined by comparing with ESCC cells expressing a control nonsilencing shRNA (shNC) and vehicle-treated ESCC cells, respectively, and are summarized using Venn diagrams. (b) Gene Set Enrichment Analysis of the overlapping differentially regulated genes in (a). (c) Heatmap of the differentially expressed genes in the “cell death,” “apoptosis,” and “ER stress” pathways. (d–f) Same as (a–c), except G9a was knocked down using shRNA or inhibited with UNC0642. (g–i) Same as (a–c), except both LSD1 and G9a were knocked down with shRNA or both LSD1 and G9a were inhibited with SP2509 and UNC0642.

Article Snippet: The following reagents were also used: hematoxylin and eosin (H&E) staining kit (Beyotime Biotech), TdT in situ apoptosis detection kit (R&D Systems, Minneapolis, MN), TRIzol reagent (Pufei Biotech, Shanghai, China), SYBR Green Supermix (Bimake, Houston, TX), RIPA buffer (Beyotime Biotech), protease inhibitors (Bimake), BCA protein assay kit (Beyotime Biotech), polyvinylidene fluoride (PVDF) membranes (Bio-Rad, Hercules, CA), Pierce ECL System (Thermo Scientific, Waltham, MA), EdU Kit (Ruibo Biotechnology Co., Ltd., Guangzhou, China), cell cycle staining kit (Multi Sciences Biotech, Hangzhou, China), annexin V-FITC/PI apoptosis detection kit (Multi Sciences Biotech), and PrimeScript RT reagent kit (Takara, Kyoto, Japan).

Techniques: Inhibition, RNA Sequencing, Expressing, shRNA, Control

Journal: Research

Article Title: Targeting the LSD1-G9a-ER Stress Pathway as a Novel Therapeutic Strategy for Esophageal Squamous Cell Carcinoma

doi: 10.34133/2022/9814652

Figure Lengend Snippet: Inhibiting both LSD1 and G9a in ESCC cells induces S-phase arrest and apoptosis. (a) Summary of the percentage of ESCC cells in the G1, S, or G2 phase after the indicated treatments for 2 days. SP2509 5 μ M, UNC0642 5 μ M. (b) Western blot analysis of the indicated cell cycle-associated proteins in ESCC cells treated as indicated. (c) EdU staining of ESCC cells treated with vehicle, 3 μ M SP2509, 1.2 μ M UNC0642, or both for 2 days; the nuclei were counterstained with Hoechst 33342. (d) Apoptosis analysis of ESCC cells after the indicated treatments for 2 days. (e) Western blot analysis of the indicated apoptosis-associated proteins in ESCC cells treated as indicated for 2 days. (f) Quantification of the indicated proteins measured in (e), expressed relative to vehicle-treated cells. (g) Representative images of ESCC cells treated with vehicle, 10 μ M SP2509, 10 μ M UNC0642, or both for 2 days. (h–k) Representative transmission electron microscopy images of ESCC cells treated as in (g); mitochondria (M) and endoplasmic reticulum (ER) are indicated. The scale bars are 5 μ m in (h), 2 μ m in (i–k). ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 (unpaired Student's t -test).

Article Snippet: The following reagents were also used: hematoxylin and eosin (H&E) staining kit (Beyotime Biotech), TdT in situ apoptosis detection kit (R&D Systems, Minneapolis, MN), TRIzol reagent (Pufei Biotech, Shanghai, China), SYBR Green Supermix (Bimake, Houston, TX), RIPA buffer (Beyotime Biotech), protease inhibitors (Bimake), BCA protein assay kit (Beyotime Biotech), polyvinylidene fluoride (PVDF) membranes (Bio-Rad, Hercules, CA), Pierce ECL System (Thermo Scientific, Waltham, MA), EdU Kit (Ruibo Biotechnology Co., Ltd., Guangzhou, China), cell cycle staining kit (Multi Sciences Biotech, Hangzhou, China), annexin V-FITC/PI apoptosis detection kit (Multi Sciences Biotech), and PrimeScript RT reagent kit (Takara, Kyoto, Japan).

Techniques: Western Blot, Staining, Transmission Assay, Electron Microscopy

Journal: Journal of Neuroscience

Article Title: Long-Latency Reductions in Gamma Power Predict Hemodynamic Changes That Underlie the Negative BOLD Signal

doi: 10.1523/jneurosci.2339-14.2015

Figure Lengend Snippet: Figure5. Spatial–temporalrepresentationofhemodynamicchangesevokedbysingle-impulsestimulation,sortedbygammapower(30–80Hz)fromtheuppercorticallayers(channels3–8) ofthesurroundelectrode(300–2000msafterstimuluspresentation).A,Hemodynamicresponseinsurroundregion,evokedbysingle-impulsestimulationofthewhiskerpad,averageofalltrials. B,Hemodynamicresponseinsurroundregion,averageoftrials(33%)havingtheleastgammapowerinthesurroundregion,intheuppercorticallayers(channels3–8).C,Hemodynamic response in surround region, average of trials (33%) having the most gamma power in the surround region, in the upper cortical layers. D, In vivo grayscale image of thinned cranial window from a single representative subject, with regions of interest around the two recording electrodes used for time-series generation. E–G, Averaged Hbt changes, evoked by single-impulse stimulation from a single representative subject, all trials (100%), least gamma (33%) and most gamma (33%) respectively, and gamma power recorded from surround electrode. Hbt changes in the whisker region response (black) and surround region response (green). Montage of images shows spatial maps of micromolar changes in Hbt. Each is averaged over a second and represents a corresponding time point in the stimulation trial. Increases in Hbt are toward the red color range, with blue representing negative changes. H, In vivo grayscale camera image of thinned cranial window from a single representative subject, with a series of concentric rings centered on the whisker barrel electrode. These were used toselectpixelsforselectionofdataasafunctionofdistanceawayfromtheelectrode.I–K,Hbtchanges,withdatafromalltrials,leastgammapower(33%)andmostgammapower(33%) respectively. The y-axis represents distance away from the tip of the whisker electrode, with each row in the image generated from pixels within each concentric ring. Stimulation represented with arrows. Error bars represent SEM across subjects. Responses averaged across subjects (n 10), except for single subject (D–H).

Article Snippet: After piercing the dura with a 27 gauge needle, a stereotaxic arm (Kopf Instruments) was used to insert the 16- channel silicone linear array electrodes (100 m spacing; area of each site, 177 m 2; Neuronexus Technologies) normal to the cortical surface to a depth of 1600 m. The probes were coupled to a preamplifier that was in turn connected to a data acquisition unit via fiber optic cable (Medusa Bioamp, Tucker Davis Technologies).

Techniques: In Vivo, Whisker Assay, Generated