Figure 7

ICSM18 inhibits prion-induced synaptotoxicity. Hippocampal neurons were treated with ICSM18 antibody and its isotype control, BRIC222 for 1.5 h before control or RML-infected brain homogenate (BH) was added to a final concentration of 0.01% w/v. BH and antibodies were then co-incubated for a further 72 h before fixation and staining. (a) Representative high-throughput images of cells stained with MAP2, spinophilin and NeuN to visualise dendrites, dendritic spines and neuronal nuclei respectively. (b) Dendritic spine image analysis mask (blue). (c,d) The white rectangles in (a) and (b) correspond to magnified views in (c) and (d) respectively. Dendritic spine density analysis for cells incubated with ICSM18 and BRIC222 before (e) RML-infected or (f) control brain homogenate treatment; The baseline was removed on a per plate basis and then normalised between 0 and 100% on a batch basis in order to run non-linear regression and shown as mean ± SEM; n = 5 independent tests; non-linear regression analysis used the equation Y = 100/\({1} + {1}0^{{(({\text{LogEC}}_{{{5}0}} \, - \,{\text{X}})\, \times \,{\text{HillSlope}})}}\) and assumed antibody-negative RML treatment group as 0.05 nM.