Fig. 3

ciMSC-EVs protect from HI-induced neuronal loss and increase cellular proliferation in the neurogenic subventricular zone. Neuronal density was analyzed via immunohistochemistry for NeuN (a) in 16-day-old C57BL/6 mice that were exposed to HI on postnatal day 9 followed by i.n. delivery of 0.9% NaCl (vehicle, Veh) or ciMSC-EVs 1, 3, and 5 days after HI. Immunohistochemistry analyses for NeuN were performed in cortex (b) and striatum (c). From the same animals, protein lysates from the entire hemisphere derived from 160-µm-thick tissue sections at the level of striatum were prepared and analyzed via western blot (d). Data were normalized to the reference protein GAPDH and to sham animals (d). The number of proliferating cells was determined via immunohistochemistry for the proliferation marker Ki67 in the subventricular zone (SVZ, e, f). Representative images in a are derived from the striatum (scale bar: 100 µm). Low magnification images in e show the total SVZ (scale bar: 200 µm); insets reveal higher magnification pictures (scale bar: 50 µm) of rectangles depicted in low magnification images. Representative western blot images in d show examples of protein abundance in tissue lysates obtained from sham-operated, HI-injured vehicle-treated, and HI-injured ciMSC-EV-treated animals. Images were cropped and scaled for illustration purposes from original western blot images provided in Suppl. Figure 6. *p < 0.05, **p < 0.01, ***p < 0.001, n = 8–10/group