Journal: iScience

Article Title: Cell-type specific circadian transcription factor BMAL1 roles in excitotoxic hippocampal lesions to enhance neurogenesis

doi: 10.1016/j.isci.2024.108829

Figure Lengend Snippet:

Article Snippet: Phospho-CREB (Ser133) (87G3) Rabbit mAb , Cell Signaling , Cat#14001; RRID: AB_2798359.

Techniques: TUNEL Assay, Quantitative RT-PCR, Software

Journal: The Journal of biological chemistry

Article Title: Characterization of an alternative splice variant of LKB1.

doi: 10.1074/jbc.M806153200

Figure Lengend Snippet: FIGURE 5. The effect of forskolin treatment on LKB1 and AMPK activity. CCL13 cells were transiently transfected with FLAG-tagged human LKB1L, LKB1S, or catalytically inactive LKB1 harboring a D194A mutation (LKB1D194A). Prior to lysis, the cells were treated in the presence or absence of 20 M forskolin for 30 min, or 0.5 mM H2O2 for 5 min. A, a nuclear enriched fraction (20–50g)wasblottedwithanti-phospho-CREB(Ser-133),orcelllysates(50– 100 g) were blotted with anti-phospho-LKB1 (Ser-428) or anti-FLAG. The blots shown are representative of blots obtained from three independent experiments. B, LKB1 activity in anti-FLAG immune complexes isolated from 50 g of total lysate protein was determined by activation of recombinant AMPK. The results are the means S.E. of three independent experiments and are plotted as units/mg total lysate protein where 1 unit is the activity of LKB1 required to activate recombinant AMPK by 1 nmol/min/mg. C, endoge- nous AMPK was immunoprecipitated from 50 g of total protein using an anti-pan antibody bound to protein A-Sepharose. AMPK activity in the immune complexes was determined using the SAMS peptide assay. The results are plotted as pmol/min/mg and are the means S.E. of three inde- pendent experiments.

Article Snippet: Mouse monoclonal antibody recognizing both LKB1L and LKB1S (Ley37D/G6) and anti-STRAD (STRAD N13) were from Santa Cruz Biotechnology.Mousemonoclonal antibody recognizing Ser(P)-428 in human LKB1L, monoclonal anti-MO25, and phospho-CREB (Ser-133, 87G3) were from Cell Signaling Technology.

Techniques: Activity Assay, Transfection, Mutagenesis, Lysis, Isolation, Activation Assay, Recombinant, Immunoprecipitation

Journal: The Journal of biological chemistry

Article Title: Characterization of an alternative splice variant of LKB1.

doi: 10.1074/jbc.M806153200

Figure Lengend Snippet: FIGURE 7. Subcellular Localization of LKB1L and LKB1S. A, CCL13 cells were transiently transfected with FLAG-tagged human LKB1L or LKB1S alone (left panels) or co-transfected with STRAD and MO25 constructs (right panels). The cells were fixed in 4% paraformaldehyde and stained with a mouse monoclonal anti-LKB1 antibody (Ley37D/G6) that recognizes both LKB1L and LKB1S. Primary antibodies were detected with fluorescently linked secondary antibodies and visualized using a Leica TCS SP1 confocal microscope. B, HEK293 cells were treated with or without 20 M forskolin for 30 min and fractionated into cytosolic (C), membrane (M), and nuclear (N) enriched frac- tions. An equal volume of each fraction was blotted with an anti-LKB1 anti- body(Ley37D/G6).Thesamefractionswereusedtodeterminetheexpression of marker proteins for each of the fractions (Cytosolic, glyceraldehydes-3- phosphate dehydrogenase (GAPDH); Membrane, Na/K ATPase; Nuclear, CREB). C, cytosolic, membrane and nuclear fractions obtained from mouse testis were blotted with an anti-LKB1 antibody (Ley37D/G6). In each case, the blots shown are representative of blots obtained from three independent experiments. The migration of molecular mass markers is indicated.

Article Snippet: Mouse monoclonal antibody recognizing both LKB1L and LKB1S (Ley37D/G6) and anti-STRAD (STRAD N13) were from Santa Cruz Biotechnology.Mousemonoclonal antibody recognizing Ser(P)-428 in human LKB1L, monoclonal anti-MO25, and phospho-CREB (Ser-133, 87G3) were from Cell Signaling Technology.

Techniques: Transfection, Construct, Staining, Microscopy, Membrane, Marker, Migration