Fig. 5. Effect of Sema3A on migration of dorsal root ganglia neurons and sympathetic neurons in vitro. (A-D) Co-cultures of the dorsal root ganglia explants from the wild-type (A,B) and homozygous neuropilin 1 mutant (C,D) embryos at E12.5 with the control HEK293T cells (A,C) and the Sema3A-secreting HEK293T cells (B,D) (2,000 cells/cm² each). (E) A cultured sympathetic ganglion neuron, immunostained with anti-neuropilin 1 antibody (green) and anti-TH antibody (originally red but yellow after merging). (F-I) Co-culture of the sympathetic ganglion explants from the wild-type (F,G) and homozygous neuropilin 1 mutant (H,I) embryos at E12.5 with the control HEK293T cells (F,H) and the Sema3A-secreting HEK293T cells (G,I) (2,000 cells/cm² each). Cultures were immunostained with anti-TH antibody. (J-M) Co-culture of the sympathetic ganglion explants from the wild-type embryos at E12.5 with the control HEK293T cells (J,L) or the Sema3A-secreting HEK293T cells (K,M) (800 cell/cm² each). The cultures were immunostained with anti-TH antibody (red) and 2H3 (green). J and K are phase-contrast micrographs. L and M correspond to the boxed regions in J and K, respectively. Arrows in A-D, J and K indicate the HEK293T cells. Scale bar: 100 µm (A-D); 25 µm (E); 200 µm (F-I); 200 µm (J,K); 200 µm (L,M).