Supplemental Fig. 2. Gradients of gap junction (GJ) pathology in MS cortex. Fol- lowing identification of lesions by MOG staining (A-C), the formation of GJ plaques by Cx30 and Cx43 in astrocytes and by Cx47 in oligodendrocytes was examined in adjacent sections in defined areas of layers 1-2 (L1), 3-4 (L3), and 5-6 (L5) of control and MS normal-appearing cortex (NAGM), and in different lesion types including subpial (SP, A), intracorti- cal (IC, B) and leukocortical lesions span- ning grey (GM) and white matter (WM) (C). Open arrowheads in A-B indicate pial surface; dashed lines outline the lesions. D-F: Representative immunostaining of Cx30/NeuN, Cx43, and Cx47 (connexins, green; NeuN, red; DAPI, blue) in different layers as indicated from control (D), NAGM (E) and different lesions (F). Open arrowheads in D-F indicate Cx47 in oligodendrocytes (asterisks in insets in D). Scale bars: A-C, 200 µm; D-F, 50 µm; insets, 10 µm. G-I: Counts of GJ plaques formed by Cx30 (G), Cx43 (H) and Cx47 (I) in different cortical layers (L1, L3, L5) of control cortex, MS NAGM, and in differ- ent types of cortical lesions (SP, IC and LC). Only significant differences (Student’s t-test) are shown with p values. Cx30 GJs are increased in subpial lesions vs. L1 of both control and MS NAGM; only in control cortex Cx30 GJs are more numerous in L3 vs. L1. Cx43 shows significantly elevated GJ numbers in both NAGM L1 and in SP lesions, vs. control L1. Cx47 GJ numbers are similar in control and MS NAGM, but are reduced in all types of cortical lesions vs. corresponding control cortex layers.