Journal: Cell Reports Medicine

Article Title: LTA4H improves the tumor microenvironment and prevents HCC progression via targeting the HNRNPA1/LTBP1/TGF-β axis

doi: 10.1016/j.xcrm.2025.102000

Figure Lengend Snippet:

Article Snippet: Anti-Histone H3 , Bioss , BSM-33042M.

Techniques: Microarray, Recombinant, Lysis, Protease Inhibitor, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Reverse Transcription, Activity Assay, Immunoprecipitation, Extraction, Purification, In Vitro, Transfection, Sequencing, Mass Cytometry, ChIP-sequencing, Transgenic Assay, Software

Journal: The Journal of Neuroscience

Article Title: GABA A Receptor Phospho-Dependent Modulation Is Regulated by Phospholipase C-Related Inactive Protein Type 1, a Novel Protein Phosphatase 1 Anchoring Protein

doi: 10.1523/JNEUROSCI.1323-04.2004

Figure Lengend Snippet: Phosphorylation of the β3 subunits of GABAA receptor by forskolin in wild-type and PRIP-1-/- mice. A, Hippocampal slices from wild-type mice were labeled with 1.5 mCi [32P]orthophosphate for 60 min and treated with 50 μm forskolin (+F) or vehicle only (-F). Homogenates (∼0.8 mg of hippocampal protein from two slices) were immunoprecipitated by anti-β2/3 antibodies (6.0 μg; clone 62-3G1; Upstate Biotechnology) or control IgG and protein G-Sepharose, followed by SDS-PAGE and autoradiography. B, Hippocampal slices from wild-type or PRIP-1-/- mice were labeled with [32P]orthophosphate and immunoprecipitated as described in A; a typical gel is shown (top) quantitated (bottom). Statistical analyses were performed using Student's t test (*p < 0.05, **p < 0.01; n = 6). C, Hippocampal slices were stimulated with forskolin (+F) for 5 min, and the extracts were analyzed by SDS-PAGE and immunoblotting. Typical immunoblots of hippocampal extracts by anti-phospho-S408/9 (P-β3) and the β2/3 subunit antibody subunit antibodies are shown in the top panels. The bottom panel shows a quantification of the blots using the phospho-specific antibody against the GABAA receptor β2/3 subunit. Results were expressed as the densities of anti-phospho-S408/9 relative to those of anti-β3. *p < 0.05 and **p < 0.01 using Student's t test (n = 4).

Article Snippet: For the assay of phospho-S408/9 of β3 subunit, hippocampal slices (400 μm thick) from wild-type and PRIP-1 -/- mice were extracted with Buffer A without BSA, followed by an SDS-PAGE and immunoblotting with antibodies specific to anti-phospho-S408/9 of β3 subunit (Brandon et al., 2002b , 2003 ; Jovanovic et al., 2004 ) or to anti-β2/3 subunit (clone 62-3G1; Upstate Biotechnology, Lake Placid, NY).

Techniques: Labeling, Immunoprecipitation, SDS Page, Autoradiography, Western Blot

Journal: The Journal of Neuroscience

Article Title: GABA A Receptor Phospho-Dependent Modulation Is Regulated by Phospholipase C-Related Inactive Protein Type 1, a Novel Protein Phosphatase 1 Anchoring Protein

doi: 10.1523/JNEUROSCI.1323-04.2004

Figure Lengend Snippet: Effect of calyculin A and okadaic acid on the phosphorylation of the GABAA receptor β3 subunits. A, Hippocampal slices labeled with 1.5 mCi [32P]orthophosphate for 60 min were incubated for 5 min in the absence or presence of forskolin (50 μm) or forskolin plus 100 nm calyculin A. Receptors were immunoprecipitated by anti-β2/3 antibodies, followed by SDS-PAGE and autoradiography, as described in Figure 1. The experiments using the slices from the wild-type and mutant mice were independently performed; therefore, each control level of the phosphorylation was taken as 100% resulting from the labeling efficiency, and results are expressed as mean ± SE from four separate experiments. *p < 0.05 and **p < 0.01 by Student's t test. B, C, Hippocampal slices were stimulated with 50 μm forskolin (+F), or together with 100 nm calyculin A (B), or 1 μm okadaic acid (C) for 5 min, and the extracts were analyzed by SDS-PAGE and immunoblotting. Higher concentrations of okadaic acid (e.g., 10 μm) could not be used in the assay systems because of toxicity. Typical immunoblots of hippocampal extracts by anti-phospho-S408/9 of the β3 subunit antibodies (top panel) and anti-β2/3 subunit antibodies (bottom panel) are shown. The graph shows the summary of the results (means ± SE) from three independent preparations. Results were expressed as the densities of anti-phospho-S408/9 relative to those of the anti-β2/3 antibody. *p < 0.05 and **p < 0.01 using Student's t test.

Article Snippet: For the assay of phospho-S408/9 of β3 subunit, hippocampal slices (400 μm thick) from wild-type and PRIP-1 -/- mice were extracted with Buffer A without BSA, followed by an SDS-PAGE and immunoblotting with antibodies specific to anti-phospho-S408/9 of β3 subunit (Brandon et al., 2002b , 2003 ; Jovanovic et al., 2004 ) or to anti-β2/3 subunit (clone 62-3G1; Upstate Biotechnology, Lake Placid, NY).

Techniques: Labeling, Incubation, Immunoprecipitation, SDS Page, Autoradiography, Mutagenesis, Western Blot

Journal: The Journal of Neuroscience

Article Title: GABA A Receptor Phospho-Dependent Modulation Is Regulated by Phospholipase C-Related Inactive Protein Type 1, a Novel Protein Phosphatase 1 Anchoring Protein

doi: 10.1523/JNEUROSCI.1323-04.2004

Figure Lengend Snippet: PRIP-1 plays a critical role in GABAA receptor phospho-dependent functional modulation regulated by D1 receptors. A, Hippocampal slices were stimulated for 5 min with SKF81297 (1 μm) together with dopamine uptake inhibitor nomifensine (10 μm). After cell lysis, the extracts were analyzed by SDS-PAGE and immunoblotting with a phospho-specific antibody against S408/9 (P-β3) in the β3 subunit or total β3 (β3), as shown in the top panels. The graph shows the summary of the results (means ± SE) from four independent experiments. Results were expressed as the densities of anti-phospho-S408/9 relative to those of anti-β2/3 antibody. *p < 0.05 by Student's t test. B, Membrane currents (IGABA) activated by application of GABA (5 μm) were recorded from mechanically dissociated hippocampal CA3 neurons. Traces represent typical GABAA receptor-mediated currents recorded at 4 min after drug application. SCH23390 (3 μm) or H-89 (3 μm) was added together with SKF81297 (1 μm). *p < 0.05 using Student's t test. C, Possible role of PRIP-1 in phospho-regulation of GABAA receptors. Neurotransmitters such as dopamine, which increases intracellular cAMP, cause the activation of PKA. This will result in enhanced phosphorylation of the GABAA receptor β1 and β3 subunits, because PKA is specifically targeted to GABAA receptors via an association with AKAP150 (Brandon et al., 2003). This enhanced phosphorylation results in functional modulation of GABAA receptors, the nature of which is subtype dependent. Activated PKA at the same time phosphorylates PRIP-1 at residues, including T94, inducing the release of active PP1cα enhancing the dephosphorylation of receptor β-subunits and hence terminating receptor functional modulation (see Results).

Article Snippet: For the assay of phospho-S408/9 of β3 subunit, hippocampal slices (400 μm thick) from wild-type and PRIP-1 -/- mice were extracted with Buffer A without BSA, followed by an SDS-PAGE and immunoblotting with antibodies specific to anti-phospho-S408/9 of β3 subunit (Brandon et al., 2002b , 2003 ; Jovanovic et al., 2004 ) or to anti-β2/3 subunit (clone 62-3G1; Upstate Biotechnology, Lake Placid, NY).

Techniques: Functional Assay, Lysis, SDS Page, Western Blot, Activation Assay, De-Phosphorylation Assay