Figure 1: Comparison of deep layer 6 (L6) neuronal populations labeled by local injections of retrograde neuronal tracers and immunostaining for Connective Tissue Growth Factor (CTGF) or Complexin 3 (Cplx3).
A, Low (Ai-Aiii) and high (Aiv-Avi) magnification images of neurons retrogradely labeled from layers 1 and 2 (L1/2; AlexaFluor 555 Cholera toxin subunit B (CTB); magenta, Ai, Aiv) and immunostained with antibodies to CTGF (green, Aii, Av) in the primary somatosensory cortex (S1) of a postnatal day 26 (P26) mouse. The two images are overlaid (Aiii, Avi) to show double-labeled neurons. B, Summary data showing the laminar distribution of retrogradely labeled neurons and CTGF-positive cells in S1 (n = 5 mice, P26-P33; light lines: individual mice; dark line: mean). C, Summary data showing the percentage of retrogradely labeled neurons in deep L6 that are CTGF-positive and the percentage of CTGF-positive neurons that are retrogradely labeled in S1 (n = 9 mice, P26-P40). D, Low (Di-Diii) and high (Div-Dvi) magnification images of neurons retrogradely labeled from L1/2 (AlexaFluor 647 CTB; magenta, Di,Div) and immunostained with antibodies to Cplx3 (green, Dii, Dv) in S1 of a P30 mouse. The two images are overlaid (Diii,Dvi) to show double-labeled neurons. E, Summary data showing the laminar distribution of retrogradely labeled neurons and Cplx3-positive cells in S1 (n = 6 mice, P28-P30; light lines: individual mice; dark lines: mean). F, Summary data showing the percentage of retrogradely labeled neurons in deep L6 that are Cplx3-positive and the number of Cplx3-positive neurons that are retrogradely labeled in S1 (n = 6 mice, P28-P30). Scale bars: Ai-iii, Di-ii: 200 μm, Aiv-vi, Div,vi: 30 μm.