Western blot analysis of fractalkine protein expression in mouse neural retina at P5 until P60 being representative of five independent experiments.

<p>Blots were re-probed with anti-GAPDH antibody. (A). Fractalkine membrane integrated (100-kDa) and soluble (85-kDa) protein forms are present in both control (wt) and rd10 retina lysates during development at P5. (B). Accumulation of presumably intracellular (95-kDa) protein form is seen in both wt (B) and rd10 (C) developing neural retina at P10. Remarkably, no cleaved (85-kDa) protein form was associated with membrane integrated (100-kDa) form in both rd10 and wt retina lysates at P10. Both wt and rd10 retinas show lower to beneath detection level of 100-kDa form and higher level of 95-kDa form at P14 through P60 (D-G). Increased level of 85-kDa form is clearly detectable in rd10 retinas at P45 and P60 (G). Histogram showing relative percent levels of each of the three CX3CL1 protein forms in wt retina (H) and degenerating rd10 retina (I). Data are expressed as percent of densitometric arbitrary units. Values are mean ± SEM, (n = 5). In rd10 retinas, positive correlation between the relative levels of a ∼100-kDa and corresponding 85-kDa bands was found at P45 and P60 (Pearson product-moment correlation coefficient r = 0.683, n = 12, p = 0.014 in RD10 retina at P45 and r = 0.882, n = 9, p = 0.0017 at P60) as well as for the relative levels of a 100-kDa and 95-kDa bands (r = 0.928, n = 11, p<0.0001 at P14, r = 0.861, n = 9, p = 0.0029 at P22, r = 0.892, n = 9, p = 0.0012 at P30, r = 0.966, n = 12, p<0.0001 at P45, and r = 0.628, n = 12, p = 0.0288 at P60 in rd10 retina). The degree of association between the levels of 95 kDa and 85 kDa protein band was r = 0.712, n = 12, p = 0.0094 at P45 and r = 0.907, n = 9, p = 0.0007 at P60. It was not, however, possible, to make similar analysis for the wild type retina samples, as the level of the ∼95-kDa protein form was the highest and the levels of cleaved and full-length forms were far below the limit of detection.