Journal: Genes, Brain, and Behavior

Article Title: Transcriptomics analysis reveals potential regulatory role of nSMase2 ( Smpd3 ) in nervous system development and function of middle‐aged mouse brains

doi: 10.1111/gbb.12911

Figure Lengend Snippet: Brain cortex deficient in nSMase2 has a different transcriptome and proteome. (A) Flow chart of the RNA‐seq experiment and data analysis. (B) The top 10 most increased transcripts and (C) decreased transcripts. The transcripts in red color were selected to be analyzed either by immunoblotting or RT‐PCR. (D) Immunoblots and quantification of protein levels of Grin2b, Map1b (antibody also detects Map1b light chain), phosphorylated Gsk3α/β (p‐Gsk3α/β) and Gapdh in fro/fro versus fro/+ cortex. (E) qRT‐PCR to confirm differences in the amounts of Naa38, Hapln2, Mrpl27, Rnaset2a, Atp5k mRNAs in fro/fro versus fro/+ cortex.

Article Snippet: Non‐specific binding sites were blocked with 5% fat‐free dry milk in PBS/0.1% Tween‐20 (PBST) followed by overnight incubation with primary antibodies, such as Map1b (Santa Cruz Biotechnology, sc‐8970), Gsk3α/β (Santa Cruz Biotechnology, sc7291), phospho‐Gsk3α/β (Santa Cruz Biotechnology, sc135653) and GAPDH (Cell signaling tech, #5174) and secondary anti‐rabbit (Jackson Immune Research, 715‐035‐152) and anti‐mouse (Jackson Immune Research, 715‐035‐151) horseradish peroxidase (HRP) antibodies for 1 h at room temperature.

Techniques: RNA Sequencing Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: eLife

Article Title: FMRP regulates postnatal neuronal migration via MAP1B

doi: 10.7554/eLife.88782

Figure Lengend Snippet: ( A ) Immunohistochemistry of the RMS showing MAP1B expression (magenta) along the stream. The RMS is delineated with dotted lines. Scale bar: 50 μm. ( B ) Immunostaining for MAP1B of a neuroblast migrating away from a ventricular/subventricular zone (V/SVZ) explant in Matrigel. The labeling is located around the nucleus and in the leading process with occasional bundles of potential microtubules (arrow). Scale bar: 10 μm.

Article Snippet: Secondary antibodies used are: ECL anti-mouse immunoglobulin G (IgG) horseradish peroxidase linked whole antibody (1/10,000; Fisher Scientific; NXA931V) for anti-MAP1B, Peroxidase-conjuguated AffiniPure F(ab’)2 Fragment Donkey Anti-Rabbit IgG (H+L) (1/5000; Jackson ImmunoResearch; 711-036-152) for anti-Vinculin.

Techniques: Immunohistochemistry, Expressing, Immunostaining, Labeling

Journal: eLife

Article Title: FMRP regulates postnatal neuronal migration via MAP1B

doi: 10.7554/eLife.88782

Figure Lengend Snippet: ( A ) Immunoblot analysis revealing the expression of MAP1B in control and Fmr1 -null micro-dissected RMS. ( B ) Quantification of MAP1B signal was performed with NIH IMAGE software and normalized to Vinculin house-keeping gene signal. MAP1B level in Fmr1 -null RMS was statistically compared and normalized to MAP1B level in control RMS (Student’s t test, p-value = 0.004) (Ctrl: N=3; MAP1B KD: N=3. **p-value <0.005.). Original and labelled files for the western blot revealing the expression of MAP1B in control and Fmr1-null micro-dissected RMS, with quantifications and statistical analysis, are available at https://osf.io/eqhzx .

Article Snippet: Secondary antibodies used are: ECL anti-mouse immunoglobulin G (IgG) horseradish peroxidase linked whole antibody (1/10,000; Fisher Scientific; NXA931V) for anti-MAP1B, Peroxidase-conjuguated AffiniPure F(ab’)2 Fragment Donkey Anti-Rabbit IgG (H+L) (1/5000; Jackson ImmunoResearch; 711-036-152) for anti-Vinculin.

Techniques: Western Blot, Expressing, Software

Journal: eLife

Article Title: FMRP regulates postnatal neuronal migration via MAP1B

doi: 10.7554/eLife.88782

Figure Lengend Snippet: ( A ) Immunohistochemistry of the RMS showing MAP1B expression (magenta) along the stream. The RMS is delineated with dotted lines. Scale bar: 50 μm. ( B ) Immunostaining for MAP1B of a neuroblast migrating away from a ventricular/subventricular zone (V/SVZ) explant in Matrigel. The labeling is located around the nucleus and in the leading process with occasional bundles of potential microtubules (arrow). Scale bar: 10 μm.

Article Snippet: Secondary antibodies used are: anti-chicken immunoglobulin Y (IgY) Alexa Fluor 488 (1/1000; Jackson ImmunoResearch; 703-545-155) against anti-GFP, anti-mouse immunoglobulin G2b (IgG2b) Alexa Fluor 555 (1/2000; Jackson ImmunoResearch; 703-545-155) against anti-FMRP and anti-MAP1B.

Techniques: Immunohistochemistry, Expressing, Immunostaining, Labeling

Journal: eLife

Article Title: FMRP regulates postnatal neuronal migration via MAP1B

doi: 10.7554/eLife.88782

Figure Lengend Snippet: ( A ) Immunoblot analysis revealing the expression of MAP1B in control and Fmr1 -null micro-dissected RMS. ( B ) Quantification of MAP1B signal was performed with NIH IMAGE software and normalized to Vinculin house-keeping gene signal. MAP1B level in Fmr1 -null RMS was statistically compared and normalized to MAP1B level in control RMS (Student’s t test, p-value = 0.004) (Ctrl: N=3; MAP1B KD: N=3. **p-value <0.005.). Original and labelled files for the western blot revealing the expression of MAP1B in control and Fmr1-null micro-dissected RMS, with quantifications and statistical analysis, are available at https://osf.io/eqhzx .

Article Snippet: Secondary antibodies used are: anti-chicken immunoglobulin Y (IgY) Alexa Fluor 488 (1/1000; Jackson ImmunoResearch; 703-545-155) against anti-GFP, anti-mouse immunoglobulin G2b (IgG2b) Alexa Fluor 555 (1/2000; Jackson ImmunoResearch; 703-545-155) against anti-FMRP and anti-MAP1B.

Techniques: Western Blot, Expressing, Software