Reactive features of astrocytes in the tubers. (a) p44+ gliotic astrocytes delineate the border between gliotic and non-gliotic areas in the peripheral parts of the tuber and are segregated from GS+ protoplasmic astrocytes. Note that only astrocytes with low levels of GS expression are p44 immunopositive (arrowheads in a1’ and a1”). (b) p44 + astrocytes in transitional border area express p-S6 (arrowheads). Giant cell (double headed arrow) expresses p44 and p-S6. p-S6+ microglia (arrow) don’t show p44 immunoreactivity. (c) p44+ gliotic astrocytes express SPARC. Note that protoplasmic astrocytes are p44 and SPARC immunonegative (arrowheads). (d) SPARC+ astrocytes express p-S6. Note that the level of p-S6 expression in protoplasmic SPARC immunonegative astrocytes (arrowheads) is less than in SPARC+ astrocytes. Confocal microscopy, double immunostaining (a,b), counterstaining with Nissl; triple immunostaining (c,d). a1’, a1”, b1’, and b1”split images from enlarged inserts in a and b, respectively. c’, c”, c”’, d’, d”, and d”’ respectively split c and d images, respectively. Scale bar: 100 μm in a, 75 μm in b, c, d.