Supplementary Figure 2: Effects of whisker lesioning at P4 on cell death, axon degeneration, and cell stress in the barrel cortex circuit.

a, Immunostaining of trigeminal ganglia, which contain the neurons that innervate the whisker follicles. There is a significant increase in ATF3 (red, marker of cell stress) in NeuN-positive neurons (cyan) at 24h post whisker lesioning (deprived, bottom row). Scale bar, 50 µm. b, Quantification for ATF3 signal co-localized to NeuN in the control and deprived trigeminal ganglia. (Two-tailed Student’s t-Test, n = 4 animals; **P = 0.0012, t = 5.715, df = 6). c, Representative images from 3 animals show ATF3 signal (green) is not detected in the ventral posterior medial nucleus (VPM) of the thalamus (bottom panels; dotted line borders the VPM) nor the primary somatosensory cortex (top panels) 24 hours after whisker lesioning. Scale bars, 150 µm. d, Quantification for cell death marker cleaved caspase 3 shows cell death does not occur 24h nor 5d after whisker lesioning in the trigeminal ganglia. (Two-way ANOVA with Sidak’s post hoc, 24h control vs deprived, n = 3 animals, P >0.9999, t = 0, df = 8; 6d control vs deprived, n = 3 animals, P = 0.3520, t = 1.414, df = 8). Data presented as mean ± SEM. e-f, Representative images from 3 animals for the deprived somatosensory cortex and VPM 7 days after whisker lesioning in control mice shows no increased cell death (e; caspase 3, green) nor axon degeneration (f; APP, green) in either brain region. Scale bars, 150 µm.