Fig. 2
From: Tissue and cellular rigidity and mechanosensitive signaling activation in Alexander disease
Increased expression of A-type laminin Alexander disease. a Double label immunofluorescence shows increased A-type lamin, lamin A/C (LA/C) expression in the astrocytes of a 1-year-old Alexander disease patient (AxD, arrows) compared to an age-matched control (Ctrl). GFAP labels astrocytes. Scale bar is 5 microns. b Western blots confirm a significant increase of lamin A expression in the white matter of Alexander disease patients compared to age-matched controls. The blots are reprobed for GAPDH to illustrate equivalent protein loading. p = 0.0476, Mann–Whitney test. c Double label immunofluorescence shows increased lamin A/C (LA/C) expression in the astrocytes of 3-month-old Alexander disease model mice (GFAPR236H/+, arrows) compared to age-matched wild-type littermate control (wild type). GFAP labels astrocytes. Scale bar is 10 microns. d Quantification of immunofluorescence intensity shows marked increase of lamin A/C expression in 3-month-old Alexander disease model mice (GFAPR236H/+) compared to age-matched wild-type mice (WT). N = 3 animals per genotype. A total of 30 astrocytes per animal were used for quantification. p = 0.0126, Wilcoxon test. e Double label immunofluorescence demonstrates increased expression of LamC, the Drosophila homolog of A-type lamin, in the glial cells of Alexander disease model flies (GFAPR79H, arrows) compared to age-matched control flies (Ctrl, arrowheads). Repo marks glial cells. Scale bar is 5 microns. f Western blot demonstrates increased expression of LamC in Alexander disease model flies compared to age-matched controls. The blot is reprobed for actin to illustrate equivalent protein loading. N = 8, p = 0.0002, Mann–Whitney test. g Reducing LamC expression using two loss-of-function alleles of LamC in Alexander disease model flies markedly reduced cell death measured by TUNEL analysis. N = 6 per genotype. *p < 0.05, **p < 0.01, Kruskal–Wallis test. h Reducing LamC expression using a loss-of-function allele of LamC in Alexander disease model flies significantly decreased the number of flies with seizures. N > 100 per genotype. p < 0.001. χ2-test. i Reducing LamC expression using a loss-of-function allele of LamC in Alexander disease model flies rescues non-cell autonomous neuronal cell death. N = 6 per genotype. p = 0.0152, Mann–Whitney test. Flies are 20 days old in e–g, i and 1 day old in h
