Fig. 5
Relative Membrane-Association of WTαA- and αAN101D in Lenses of CRYαAN101D and CRYαAWT Mice. a, b, e and f: the relative levels of association of WTαA and αAN101D with the purified membrane preparations at different membrane purification steps analyzed using SDS-PAGE analysis at two different age groups (1 and 6 months) (c, d, g and h): Similarly the samples were analyzed by Western blot using anti-His antibody at two different age groups (1and 6 months). Additionally, in both left and right panels, the lanes 1, 2 and 3 show the WS-protein fractions recovered after 1st, 2nd and 3rd consecutive washes in buffer B to solubilize WS-proteins, respectively. Lanes 4 and 5 represent the urea soluble-protein fractions recovered during two consecutive washes of WI-protein pellet with buffer B containing 8 M urea, similarly, lanes 7 and 8 from 6-month old lenses (E and F) represent purified membrane preparation. Lane 9 of 6-month old lenses represents crude WS-homogenate. i & j: Quantification of immunoreactive bands of αA- recovered in urea-soluble fractions (Lane 4 [L4] and lane 5 [L5] represent two consecutive urea wash of WI proteins during membrane isolation from lenses of CRYαAWT and CRYαAN101Dmice as shown in Western blot analysis in Fig. 5. i Relative levels of immunoreactive WTαA lenses (blue) and αA-N101D αA- (red) during membrane purification from 1-month old lenses. j Relative levels of immunoreactive WTαA lenses (blue) and αAN101D αA- (red) during membrane purification from 6-months old lenses. Note that relatively higher levels of αAN101D than WTαA was associated with purified membranes in lanes 4 (in 1-month old) and lane 5 (in both 1- and 6- month-old) of the two types of lenses. Gel images are not cropped