Extended Data Fig. 9: Additional experiments investigating OPN receptors and OPN KD.
From: Single-cell and spatial RNA sequencing identify perturbators of microglial functions with aging
a) Lesions of 6wk and 52wk mice 7 days after PAzePC injection labeled with CD68 (red) and ITGB1 (green) (left), IBA1 (red) and ITGAV (green) (middle), and CD68 (red) and ITGB5 (green) (right). b-d) Graphs comparing the percent of ITGB1 (b), ITGAV (c), and ITGB5 (d) found in the PAzePC lesions of 6wk and 52wk mice. Data from 2 separate experiment, n =6 per experimental group. Significance indicated as *** p < 0.001, two-tailed, unpaired t-test, comparing 6wk and 52wk. Data are represented as mean ± SD. e) Lesions of 52wk mice injected with Ctrl or OPN KD AAV 7 days after PAzePC treatment labeled for CD68 (red) and GFP (green) (left), NFH (red) and GFP (green) (middle), and GFAP (red) and GFP (green) (right). f) Graph comparing the expression of GFP in CD68+ microglia/macrophage, NFH+ axons, and GFAP+ astrocytes from Ctrl or KD treated spinal cords. g) Lesions of 52wk mice injected with Ctrl or OPN KD AAV 7 days after PAzePC treatment labeled for CD68 (green) and MMP9 (red) (left) or CD68 (red) and MMP14 (red) (right). h) Graphs comparing the percent of MMP9 (left) and MMP14 (right) found in the PAzePC lesions of Ctrl and OPN KD mice. Data were acquired from 2 separate experiments, n = 6 per experimental group. Significance indicated as * p < 0.05, unpaired t-test, comparing Ctrl and OPN KD treated 52wk mice after PAzePC injection. Data are represented as mean ± SD. i) Lesions of 52wk mice injected with AAV6 with a CD68 promoter to express scrambled shRNA (CD68 Ctrl) or OPN targeting shRNA (CD68 OPN KD) 7 days after PAzePC treatment labeled with E06 (green), CD16/32 (red), NFH (grey) (left), or with GFP (green) and CD68 (red) (middle left), or with GFP (green) and NFH (red) (middle right), or with GFP (green) and GFAP (right). j) Graph comparing NFH+ axon density in the PAzePC lesions from CD68 Ctrl and CD68 OPN KD treated 52wk mice. k) Graph comparing the expression of GFP in CD68+ microglia/macrophage, NFH+ axons, and GFAP+ astrocytes from CD68 Ctrl or CD68 OPN KD treated spinal cords. Data were acquired from 2 separate experiments, n = 6 per experimental group. Significance indicated as * p < 0.05, unpaired t-test, comparing CD68 Ctrl and CD68 OPN KD treated 52wk mice after PAzePC injection. Data are represented as mean ± SD.
