Scaffold-free, size-controlled generation of spheroids for biochemical assays, drug screening and high-content imaging:Protocol

This protocol will allow the user to establish size-controlled, scaffold-free spheroid cultures suitable for drug screening, biochemical signalling assays and high-content imaging. This protocol has been tested on and worked successfully with the following cell lines: HeLa, MCF10A, T47D, BT474

Cellular protein extraction and Western blotting using dry transfer (iBlot system):Protocol

A generic protein extraction and Western blotting protocol, allowing both for total cell lysis or cytoplasmic protein extraction only. The cytoplasmic extraction version has been used when Western blotting for PI3K and MAPK/ERK signalling components; total cell lysis with sonication has been used when Western blotting for histones

Slides given as part of the “Improving the research process: discussing an ‘open research’ policy” event held on the 8th of June 2016.

These slides were presented by Ralitsa Madsen during the “Improving the research process: discussing an ‘open research’ policy” event held by the Office of Scholarly Communication on the 8th of June 2016. The write-up of the event can be found here: https://www.repository.cam.ac.uk/handle/1810/256838

SIGNAL-seq:Multimodal Single-cell Inter- and Intra-cellular Signalling Analysis

We present SIGNAL-seq (Split-pool Indexing siG-Nalling AnaLysis by sequencing): a multiplexed splitpool combinatorial barcoding method that simultaneously measures RNA and post-translational modifications (PTMs) in fixed single cells from 3D models. SIGNAL-seq PTM measurements are equivalent to mass cytometry and RNA gene detection is analogous to split-pool barcoding scRNA-seq. By measuring both mRNA ligand-receptor pairs and PTMs in single cells, SIGNAL-seq can simultaneously uncover inter- and intra-cellular regulation of tumour microenvironment plasticity