Fig 1.
NS-Pten KO leads to increased expression of FMRP.
Hippocampal tissue from NS-Pten wildtype (WT) and knockout (KO) mice were examined for total FMRP levels using western blotting. (Fig 1A) The figure shows a representative blot from two NS-Pten WT and KO samples. (Fig 1B) Graphs show the mean (± SEM) of WT and KO mice. *** = p < 0.001. n = 9 per group.
Fig 2.
NS-Pten KO leads to increased expression of S499-Phosphorylated FMRP.
(Fig 2A) S499-phosphorylated FMRP expression indicated by the antibody from PhosphoSolutions® and abcam® for NS-Pten wildtype (WT) and knockout (KO) mice. The figures are representative blots from WT and KO hippocampal samples probed for the respective antibodies. (Fig 2B) S499-phosphorylated FMRP expression indicated by the Phosphosolutions® antibody. (Fig 2C) S499-phosphorylated FMRP expression indicated by abcam®’s antibody NS-Pten for WT and KO mice. Graphs show the mean (± SEM). n = 9 per group. ** = p < 0.01; *** = p < 0.001.
Fig 3.
Comparison of S499-Phosphorylated FMRP Expression Between Antibodies.
(Fig 3A) Comparison of S499-phosphorylated FMRP expression indicated between the antibodies from PhosphoSolutions® and abcam® in NS-Pten WT mice (Fig 3B) Comparison of S499-phosphorylated FMRP expression indicated between PhosphoSolutions® and abcam® antibodies in NS-Pten KO mice. ** = p < 0.01. Graphs show the mean (± SEM). n = 9 per group.
Fig 4.
Comparison of S499-Phosphorylated FMRP Expression Between Antibodies.
(Fig 4A) Correlation of S499-phosphorylated FMRP expression indicated between PhosphoSolutions® and abcam® antibodies in NS-Pten WT mice. (Fig 4B) Correlation of S499-phosphorylated FMRP expression indicated between PhosphoSolutions® and abcam® antibodies in NS-Pten KO mice. The line represents the line of best fit. n = 9 per graph.
Fig 5.
Comparison of S499-Phosphorylated FMRP Expression Between Antibodies.
We examined phosphorylated levels of FMRP in Fmr-1 wildtype (WT) and knockout (KO) mice using 4 concentrations of the abcam® and PhosphoSolutions® antibody. The full blot of the samples is presented in each set of figures. There are 2 alternating lanes of WT and KO probed samples throughout all dilutions. In each dilution experiment the first figure used the abcam® S499 phosphorylated antibody (i) and the second figure used the PhosphoSolutions® S499 phosphorylated antibody (ii). The blue arrows point to the location of the target site. (Fig 5A) WT and KO hippocampus tissue probed for antibodies using a 1:250 dilution, (Fig 5B) 1:500 dilution, (Fig 5C) 1:1000 dilution, (Fig 5D) 1:200 dilution