Journal: Oncology Letters

Article Title: Increased soluble E‑cadherin of spheroid formation supplemented with fetal bovine serum in colorectal cancer cells

doi: 10.3892/ol.2023.13793

Figure Lengend Snippet: Densitometric results of western blotting on SNU-C5 and SNU-C5/5-FUR cells.

Article Snippet: CREB (1:1,000; cat. no. CSB-PA005947HA01HU; Cusabio Technology, LLC), fibronectin (1:2,000; cat. no. CL54951AP; Cedarlane Laboratories), p21 (1:1,000; cat. no. 60214-1; Proteintech Group, Inc.) and p90RSK (Ab348; 1:1,000; cat. no. 79-554; Prosci, Inc.) were obtained from the corresponding listed company.

Techniques: Western Blot, Marker

Journal: eLife

Article Title: Pathogenic PS1 phosphorylation at Ser367

doi: 10.7554/eLife.19720

Figure Lengend Snippet: ( A ) FLIM analysis of the PS1 conformation in 7 W cells. The cells were pre-treated with 30 µM H-89 or 1 µM KT5720 for 16 hr, followed by the treatment with vehicle control (−) or 5 µM A23187 (+) for 15 min. The FRET efficiency in each group is normalized to the average FRET efficiency of vehicle control treated neurons (n = 37–52 cells). Mean ± SEM, **p<0.01, ***p<0.001, one-way factorial ANOVA. ( B ) Western Blot analysis of PKA-dependent CREB S133 phosphorylation. Primary neurons were pre-treated with vehicle control or 1 µM KT5720 for 16 hr, followed by the treatment with vehicle control or 50 mM KCl for 5 min. Treatment with 10 µM forskolin was used as a positive control of PKA activation. ( C ) Spectral FRET assay of the PS1 conformation in primary neurons. AAV-G-PS1-R-transduced primary neurons were pre-treated with 1 µM KT5720 for 16 hr, followed by the treatment with 5 mM KCl for 5 min. Pre: before KCl application, Post: 5 min after KCl application. n = 14–17 cells, Mean ± SEM, *p<0.05, **p<0.01, ***p<0.001, one-way factorial ANOVA. ( D ) Dominant negative PKA regulatory subunit α (G324D PKA) was co-transfected with WT G-PS1-R into 7 W cells. The cells were treated with vehicle control or 5 µM A23187 for 15 min. Western Blot analysis of the PS1 S310 phosphorylation (top panel). Spectral FRET assay of the PS1 conformation (bottom panel). n = 83–88 cells, Mean ± SEM, *p<0.05, ***p<0.001, one-way factorial ANOVA. DOI: http://dx.doi.org/10.7554/eLife.19720.010

Article Snippet: Rabbit monoclonal CREB antibody (RRID: AB_309979 ) was from Upstate Biotechnology (Lake Placid, NY, USA).

Techniques: Control, Western Blot, Phospho-proteomics, Positive Control, Activation Assay, Dominant Negative Mutation, Transfection

Journal: eLife

Article Title: Pathogenic PS1 phosphorylation at Ser367

doi: 10.7554/eLife.19720

Figure Lengend Snippet: ( A ) Immunohistochemical detection of the phosphorylated CREB in mouse brain injected with 100 mM 8-Bromo-cAMP (right hemisphere) or vehicle (left hemisphere). The brain sections were immunostained with a p-CREB S133 antibody. Strong fluorescence in cAMP injected hemisphere shows PKA activation. A scale bar indicates 50 µm. Mean ± SEM, ***p<0.001, Student’s t-test. ( B ) Antibody-based spectral FRET validation. The brain sections were immunostained with PS1 NT-A488 (donor only, negative control for FRET), PS1 NT-A488-Cy3 (A488-Goat-anti-mouse IgG was detected with Cy3-Donkey-anti-goat IgG, positive control), or PS1 NT-A488/PS1 CT-Cy3 (FRET, PS1 NT-PS1 CT proximity) antibodies. The Y axis shows the ratio of fluorescence intensities within the 522 nm to 672 nm spectral range normalized by the emission intensity at 522 nm. The positive control (in green) shows large ‘hump’ at 565/522 nm, and the FRET signal in PS1 NT-PS1 CT stained cells (in red) shows smaller but significant increase in the 565 nm/522 nm ratio compared to the donor only (n = 75–179 cells). Mean ± SEM, ***p<0.001 vs. Donor only, one-way factorial ANOVA. ( C ) Immunohistochemical detection of the phosphorylated CREB S133 in mouse brain injected with 100 µM KT5720 (right hemisphere) or vehicle (left hemisphere), followed by the injection of 100 mM 8-Bromo-cAMP (both hemispheres). A scale bar indicates 50 µm. Mean ± SEM, ***p<0.001, Student’s t-test. DOI: http://dx.doi.org/10.7554/eLife.19720.015

Article Snippet: Rabbit monoclonal CREB antibody (RRID: AB_309979 ) was from Upstate Biotechnology (Lake Placid, NY, USA).

Techniques: Immunohistochemical staining, Injection, Fluorescence, Activation Assay, Biomarker Discovery, Negative Control, Positive Control, Staining

Journal: British Journal of Pharmacology

Article Title: Prenatal and postnatal alcohol exposure increases vulnerability to cocaine addiction in adult mice

doi: 10.1111/bph.14901

Figure Lengend Snippet: Primary antibodies

Article Snippet: The Immuno‐related procedures used comply with the recommendations made by the British Journal of Pharmacology (Alexander et al., 2018 ). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Primary antibody Description Host Dilution Company Catalogue number RRID ΔFosB Delta FosB Mouse 1:250 Abcam #ab11959 AB_298732 CREB cAMP‐response element binding protein Rabbit 1:500 MERCK #04‐767 AB_1586959 D 1 R Dopamine receptor 1 Rabbit 1:500 Abcam #ab20066 AB_445306 D 2 R Dopamine receptor 2 Rabbit 1:500 MERCK #324393 AB_10681601 DARPP‐32 Dopamine‐ and cAMP‐regulated phosphoprotein Rabbit 1:1,000 Cell Signaling #2306 AB_823479 GAPDH Glyceraldehyde‐3‐phosphate dehydrogenase Mouse 1:2,500 Santa Cruz Biotechnology #sc‐365062 AB_10847862 GluA1 AMPA receptor subunit 1 Rabbit 1:1,000 MERCK #ABN241 AB_2721164 GluA2 AMPA receptor subunit 2 Rabbit 1:1,000 MERCK #AB1768‐I AB_2313802 pCREB Phosphorylated CREB Rabbit 1:1,000 MERCK #06‐519 AB_310153 pDARPP‐32 (Thr34) Phosphorylated DARPP‐32 (Thr34) Rabbit 1:1,000 MERCK #ab9206 AB_347689 β‐Tubulin β‐Tubulin Mouse 1:5,000 BD PharMingen #556321 AB_396360 β‐Tubulin Class III β‐tubulin Rabbit 1:1,000 Abcam #ab6046 AB_2210370 Open in a separate window Primary antibodies

Techniques: Binding Assay

Journal: British Journal of Pharmacology

Article Title: Prenatal and postnatal alcohol exposure increases vulnerability to cocaine addiction in adult mice

doi: 10.1111/bph.14901

Figure Lengend Snippet: GluA1/GluA2 ratio and striatal ΔFosB expression were altered in PLAE mice after cocaine‐primed reinstatement session. (a–f) Western blot analyses of GluA1/GluA2, pCREB/CREB, pDARPP‐32/DARPP‐32, ΔFosB, D1R, and D2R protein expression in the PFC and STR of PLAE and control mice following cocaine‐induced reinstatement. Student's t test, * P < .05 control versus PLAE group (n = 5–6 per group). The numbers in the bars represent the amount of individuals in the group. The lower panels show representative fluorescent immunoblots

Article Snippet: The Immuno‐related procedures used comply with the recommendations made by the British Journal of Pharmacology (Alexander et al., 2018 ). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Primary antibody Description Host Dilution Company Catalogue number RRID ΔFosB Delta FosB Mouse 1:250 Abcam #ab11959 AB_298732 CREB cAMP‐response element binding protein Rabbit 1:500 MERCK #04‐767 AB_1586959 D 1 R Dopamine receptor 1 Rabbit 1:500 Abcam #ab20066 AB_445306 D 2 R Dopamine receptor 2 Rabbit 1:500 MERCK #324393 AB_10681601 DARPP‐32 Dopamine‐ and cAMP‐regulated phosphoprotein Rabbit 1:1,000 Cell Signaling #2306 AB_823479 GAPDH Glyceraldehyde‐3‐phosphate dehydrogenase Mouse 1:2,500 Santa Cruz Biotechnology #sc‐365062 AB_10847862 GluA1 AMPA receptor subunit 1 Rabbit 1:1,000 MERCK #ABN241 AB_2721164 GluA2 AMPA receptor subunit 2 Rabbit 1:1,000 MERCK #AB1768‐I AB_2313802 pCREB Phosphorylated CREB Rabbit 1:1,000 MERCK #06‐519 AB_310153 pDARPP‐32 (Thr34) Phosphorylated DARPP‐32 (Thr34) Rabbit 1:1,000 MERCK #ab9206 AB_347689 β‐Tubulin β‐Tubulin Mouse 1:5,000 BD PharMingen #556321 AB_396360 β‐Tubulin Class III β‐tubulin Rabbit 1:1,000 Abcam #ab6046 AB_2210370 Open in a separate window Primary antibodies

Techniques: Expressing, Western Blot