Expression and localisation of C5 and C5a in hSOD1G93Aand wild-type mice during disease progression. (A) mRNA expression profile of C5 in lumbar spinal cord of hSOD1G93A mice relative to wild-type (WT) mice. Dashed line, baseline expression in WT controls at each time point. (B) Protein expression of C5a in the lumbar spinal cord of hSOD1G93A mice has decreased by end stage (ES) when compared with WT mice. (A, B) Data expressed as mean ± standard error of the mean (n = 6 mice/group; *P <0.05, Student t test). (C) to (H) Double immunolabelling for C5 (red) with cellular makers (green) for motor neurons (ChAT; C and D, arrows), astrocytes (glial fibrillary acidic protein (GFAP), E and F), and microglia (CD11b; G and H), in the ventral lumbar spinal cord region of hSOD1G93A mice (D, F and H) and WT mice (C, E and G) at end stage. Co-localisation of C5 with these cellular markers is seen as a merge of green and red (for example, white arrows in C, D, and H). PS, pre-symptomatic; OS, onset; MS, mid-symptomatic; ES, end-stage. Scale bar for C to H = 20 μm.