Journal: bioRxiv

Article Title: Human pluripotent stem cell modeling of tuberous sclerosis complex reveals lineage-specific therapeutic vulnerabilities

doi: 10.1101/683359

Figure Lengend Snippet: (A) Phase images of WT and TSC2 −/− cells during monolayer-NPC induction at differentiation days 5 and 12, displaying increased cell size and vesicle accumulation in TSC2 −/− cultures. Scale bar, 50µm. (B) Exemplary western blot of H1 and H7 NPC cultures, probing for TSC2, P-S6 and S6, revealing constitutive S6 phosphorylation in TSC2 −/− cells. (C) Quantification of NPC volume, relative to WT by Cellomics ArrayScan imaging analysis. Values are the mean ± SEM (n=13, including all cell lines). (D) Quantification of PAX6 fluorescence intensity in NPCs, relative to WT NPCs. Values are mean ± SEM (n = 5; 1x H7, 2x for H1 and 168 at p0-3). (E) Quantification of SOX2 fluorescence intensity in NPCs over early, mid, and late passage ranges, normalized to WT NPCs. Values are mean ± SEM (n = 6 early, n = 5 mid, n = 3 late). (F) Representative immunostaining for neuronal marker MAP2 (top image) and glial marker GFAP (bottom image). Graph (bottom panel) displaying % of cells in neuronal cultures expressing high levels of GFAP relative to WT. (n = 4; 2x each H7 and H9). Scale bar, 50µm. (G) Representative traces of current clamp recordings of action potentials (left) and voltage clamp recordings of AMPA receptor-mediated sEPCs (top) in WT , TSC2 −/− , and TSC2 −/− +rapamycin neurons. Quantification of sEPSC frequency (bottom). Values are mean ± SEM.

Article Snippet: Imaging for high content quantification was performed using the Thermo Fisher ArrayScan VTI High Content Screening (HCS) platform, utilizing the live cell chamber at 37°C, 5% CO 2 .

Techniques: Western Blot, Phospho-proteomics, Imaging, Fluorescence, Immunostaining, Marker, Expressing

Journal: Oncotarget

Article Title: Efficacy of aerosol therapy of lung cancer correlates with EGFR paralysis induced by AvidinOX-anchored biotinylated Cetuximab

doi:

Figure Lengend Snippet: Fluorescence imaging by High Content Screening (HCS) Operetta of A549 cells, with or without AvidinOX conjugation (100 μg/mL), incubated with 5 μg/mL CF488-labelled bMabs (blue). At indicated time, cells were washed, fixed and stained for the detection of EGFR by AF555-labeled anti-EGFR Mab (D38B1) (red). Draq5 dye staining of nucleus and cytoplasm (yellow). Violet is the result of blue and red dye co-localization in the merged images. A, bCet, 2 h cultivation. B, bCet, bPan or bRit, 30 min or 30 min contact and 24 h cultivation. C, Unlabelled bCet 30 min contact and 24 h cultivation. Staining of lysosomes by LysoTracker (light blue) and staining of EGFR as before. Hoechst dye staining of nuclei (yellow). All panels: representative picture of at least 5 fields of triplicate wells. Magnification 60X.

Article Snippet: EGFR was detected by AF555-conjugated rabbit anti-EGFR (D38B1) (Cell Signaling), added after cell fixation with 4% paraformaldehyde in PBS, permeabilization with PBS 0.2%Tween-20 (PBS-T) and blocking with 2% BSA in PBS-T.

Techniques: Fluorescence, Imaging, High Content Screening, Conjugation Assay, Incubation, Staining, Labeling