Journal: Cell & bioscience

Article Title: Rasal1 regulates calcium dependent neuronal maturation by modifying microtubule dynamics.

doi: 10.1186/s13578-024-01193-w

Figure Lengend Snippet: Fig. 1 Rasal1 is expressed in the soma and neurites of developing hippocampal neurons. A Expression of Rasal1 in coronal section of adult mouse brain. Allen Mouse Brain Atlas, http://mouse.brain-map.org/experiment/show/75774111. B Western blot analyses with Rasal1 antibody in the absence (left) and presence (right) of Rasal1 immunizing peptide. Lane 1: mouse hippocampus; lane 2: mouse prefrontal cortex; lane 3: rat hippocampus all at P8. C Endogenous Rasal1 immunoreactivity on DIV2, DIV4, DIV6, DIV12 and DIV14 in neurons stained for Rasal1 (red) and Tubulin (blue) and imaged with confocal microscopy. DIV 2 scale bar: 5 μm, DIV 4–10 scale bar: 10 μm

Article Snippet: We compared several Rasal1 antibodies; Rasal1 sc-68771 Santa Cruz, Rasal1 NBP1-32776 Novus Biologicals and Rasal1 from GenScript (Piscataway, NJ).

Techniques: Expressing, Western Blot, Staining, Confocal Microscopy

Journal: Cell & bioscience

Article Title: Rasal1 regulates calcium dependent neuronal maturation by modifying microtubule dynamics.

doi: 10.1186/s13578-024-01193-w

Figure Lengend Snippet: Fig. 3 Rasal1 binds to CaMKII, PKC, α- and β-tubulin. Confirmation of suspected Rasal1 binding partners using pull-down assay (n = 3) with GST tagged Rasal1 fragments; N-terminus (GST-n), middle fragment (GST-m) or C-terminal fragment (GST-c) validated by Co-IP. A Western blot of pull-down assay (left) and Co-IP (right) of GST-tagged Rasal1 fragments stained for CaMKII in hippocampal and whole brain lysate. B Western blot of pull-down assay (left) and Co-IP (right) of GST-tagged Rasal1 fragments stained for PKC in hippocampal and whole brain lysate. C Western blot of pull-down assay (left) and Co-IP (right) of GST-tagged Rasal1 fragments stained for β-tubulin in hippocampal and whole brain lysate

Article Snippet: We compared several Rasal1 antibodies; Rasal1 sc-68771 Santa Cruz, Rasal1 NBP1-32776 Novus Biologicals and Rasal1 from GenScript (Piscataway, NJ).

Techniques: Binding Assay, Pull Down Assay, Co-Immunoprecipitation Assay, Western Blot, Staining

Journal: Cell & bioscience

Article Title: Rasal1 regulates calcium dependent neuronal maturation by modifying microtubule dynamics.

doi: 10.1186/s13578-024-01193-w

Figure Lengend Snippet: Fig. 4 Rasal1 inhibits neurite growth. A Neurite tracing using SynD [29] at DIV 6, 8, 10, 12 and 14 of mouse hippocampal neurons transduced with either Rasal1-shRNA-EGFP, Rasal1-OE, or EGFP-control. Scale bar: 50 µm. B Neurite length (µm) of EGFP-control, Rasal1-shRNA, and Rasal1-OE expressing mouse hippocampal neurons at DIV 6, 8, 10, 12, and 14. Data pooled from 4 independent experiments (n = 122). Mean ± SEM are plotted, *p < 0.05, **p < 0.01, ***p < 0.001, unpaired student t-test, two tailed

Article Snippet: We compared several Rasal1 antibodies; Rasal1 sc-68771 Santa Cruz, Rasal1 NBP1-32776 Novus Biologicals and Rasal1 from GenScript (Piscataway, NJ).

Techniques: Transduction, shRNA, Control, Expressing, Two Tailed Test

Journal: Cell & bioscience

Article Title: Rasal1 regulates calcium dependent neuronal maturation by modifying microtubule dynamics.

doi: 10.1186/s13578-024-01193-w

Figure Lengend Snippet: Fig. 5 Rasal1 inhibits dendrite length and branching. A Representative confocal images of EGFP-control, Rasal1-OE, and Rasal1-shRNA transduced mouse hippocampal neurons stained for the dendritic marker MAP2 (red) and axonal marker Tau1 (green) (left). Tracings of dendrites (middle) and axonal neurites (right) were done using SynD. Scale bar: 20 µm. B Total axon and dendrite length (µm) of EGFP-control, Rasal1-OE, and Rasal-shRNA expressing neurons. C Number of primary and secondary dendritic branches in neurons expressing EGFP-control, Rasal1-OE, and Rasal1-shRNA. Data information: In B, C Mean ± SEM are plotted, *p < 0.05, **p < 0.01, ***p < 0.001, unpaired student t-test, two tailed. n = 60 for each group, pooled from 3 repeated experiments

Article Snippet: We compared several Rasal1 antibodies; Rasal1 sc-68771 Santa Cruz, Rasal1 NBP1-32776 Novus Biologicals and Rasal1 from GenScript (Piscataway, NJ).

Techniques: Control, shRNA, Staining, Marker, Expressing, Two Tailed Test

Journal: Cell & bioscience

Article Title: Rasal1 regulates calcium dependent neuronal maturation by modifying microtubule dynamics.

doi: 10.1186/s13578-024-01193-w

Figure Lengend Snippet: Fig. 6 Rasal1 increases tubulin detyrosination. A EGFP-control, Rasal1-OE, and Rasal1-shRNA transduced hippocampal neurons stained for tyrosinated tubulin (red) and detyrosinated tubulin (green) at DIV6. Scale bar: 20 μm unless stated otherwise. B, C Quantification of fluorescence intensity measured in the mid region of the longest neurite of detyrosinated (B) and tyrosinated (C) tubulin as a proportion of total tubulin. n = 50 for each group, pooled from 5 repeated experiments, mean plus SEM are plotted, *** = p < 0.001, unpaired student t-test, two tailed

Article Snippet: We compared several Rasal1 antibodies; Rasal1 sc-68771 Santa Cruz, Rasal1 NBP1-32776 Novus Biologicals and Rasal1 from GenScript (Piscataway, NJ).

Techniques: Control, shRNA, Staining, Fluorescence, Two Tailed Test

Journal: Cell & bioscience

Article Title: Rasal1 regulates calcium dependent neuronal maturation by modifying microtubule dynamics.

doi: 10.1186/s13578-024-01193-w

Figure Lengend Snippet: Fig. 9 Working model of proposed mechanistic activities of Rasal1 in neurites and synapses

Article Snippet: We compared several Rasal1 antibodies; Rasal1 sc-68771 Santa Cruz, Rasal1 NBP1-32776 Novus Biologicals and Rasal1 from GenScript (Piscataway, NJ).

Techniques: