Figure 7.

In vitro derived sensory neurons as a surrogate model for peripheral nerve injury. (a–c) Phase contrast images of control (a) and H₂O₂ treated (b,c) in vitro sensory neurons following 48 h culture in the absence (b) or presence (c) of the pan-caspase inhibitor Z-VAD-FMK. Exposure of the differentiated cultures to H₂O₂ induces neurite retraction (b) while inclusion of Z-VAD-FMK (c) blocks this process and the neuronal networks are largely preserved. (d) Immunoblotting of control and H₂O₂ treated in vitro sensory neuron cultures. Exposure of the cells to H₂O₂ for 24 h induces the upregulation of both CASP3 and CASP12. Also note the processed 38 kDa CASP12 fragment in two of the three cultures. ACTB was used as a loading control. The original full-length immunoblotting images are presented in Supplementary Fig. 4. (e–h) Immunocytochemical staining for CASP3 (e–f) and CASP12 (g–h) in control (e and g) and H₂O₂ treated (f and h) in vitro sensory neuron cultures. Numerous CASP3⁺ (f) and CASP12⁺ (h) cells are observed after exposure of the cultures to H₂O₂ for 24 h_. Moreover, extensive disintegration of NEFH⁺ axon bundles (f and h) is also observed upon H₂O₂ treatment. Scale bars: (a–c) 100 µm, (e–h) 40 µm.