Figure 3.

Recruitment of cytosolic chaperones to PMP22 aggresomes. Teased sciatic nerves from Wt (A, B, insets) and TrJ (A-E) mice were double immunostained with anti-PMP22 (A, C-E, red) and anti-Hsc70 (B-D, green) or anti-transferrin receptor (E, green) antibodies. Nuclei are stained with Hoechst dye (A-C, E). In A-E, arrows indicate PMP22 aggresome-like structures, whereas arrowheads point at distal PMP22-positive structures. Scale bar: A-C, E, 10 μm. A confocal image of teased TrJ nerve fiber immunostained with anti-PMP22 (D, red) and anti-Hsc70 (D, green) antibodies is shown (D). A magnification (2×) of the perinuclear region of a TrJ SC (D, asterisk) is shown in the bottom left corner (D, box). Representative planes (D₁-D₄) of the composite image shown in D illustrate the spatial relationship between PMP22 and Hsc70. Distal PMP22-containing structures (E, arrowheads) costain with transferrin receptor (E, green) but not with Hsc70 (A-C). Total (T), IPB-soluble (S), and IPB-insoluble (I) fractions of Wt and TrJ sciatic nerve lysates (40 μg/lane) were analyzed by Western blots using anti-Hsp70, anti-αB-crystallin (αBc), anti-calreticulin (CRT), and anti-GAPDH (loading control) antibodies (F). Molecular mass is in kilodaltons.