Skip to main content
. 2022 Oct 31;11(21):3448. doi: 10.3390/cells11213448

Figure 10.

Figure 10

Western blot analysis of RNA methylation-related enzymes in spinal cord and skeletal muscle of hSOD1 tg mice. (A) FTO levels in spinal cord ((A), top) and skeletal muscle ((A), bottom) of nonconditional hSOD1 tg mice (presymptomatic and symptomatic) and conditional hSOD1mus tg mice. Age-matched non-tg littermates were controls. The several different hSOD1 tg mouse lines are shown randomized among the lane assignments. FTO size (in kDa) is indicated for each blot. Ponceau S-stained membranes show protein loading. (B) METTL3 levels in spinal cord ((B), top and middle) and skeletal muscle ((B), bottom) of nonconditional hSOD1 tg mice (presymptomatic and symptomatic) and conditional hSOD1mus tg mice. Age-matched non-tg littermates were controls. METTL3 size (in kDa) is indicated. In top blot, different subcellular fractions of spinal cord were used to assess the presence of METTL3 in mitochondrial-enriched (P2), soluble (S2), and nuclear-enriched (P1) tissue compartments. The middle blot was done using nuclear fractions of spinal cord. The lowest blot in B was done using nuclear fractions of skeletal muscle. (C) ALKBH5 levels in spinal cord ((C), top and middle) and skeletal muscle ((C), bottom) of nonconditional hSOD1 tg mice (presymptomatic and symptomatic) and conditional hSOD1mus tg mice. Age-matched non-tg littermates were controls. ALKBH5 size (in kDa) is indicated. In top blot, different subcellular fractions of spinal cord were used to assess ALKBH5 presence in the mitochondrial-enriched, soluble, and nuclear-enriched compartments and crude homogenate. The middle blot was done using mitochondrial fractions of spinal cord. The lowest blot in C was done using mitochondrial fractions of skeletal muscle. (D) Protein IR levels (mean ± SD, as % of non-tg mouse immunodensity) for FTO, METTL3, and ALKBH5 in spinal cord of nonconditional hSOD1-G93A mice at presymptomatic and symptomatic stages of disease (n = 6/group). * p < 0.001; + p < 0.05; ** p < 0.01; *** p < 0.001. (E) Protein IR levels (mean ± SD, as % of non-tg mouse immunodensity) for FTO, METTL3, and ALKBH5 in skeletal muscle of nonconditional hSOD1-G93A mice at presymptomatic and symptomatic stages of disease (n = 6/group). * p < 0.001; ** p < 0.01; *** p < 0.001.