Journal: BMC Genomics

Article Title: Distinct and overlapping gene regulatory networks in BMP- and HDAC-controlled cell fate determination in the embryonic forebrain

doi: 10.1186/1471-2164-13-298

Figure Lengend Snippet: Western Blot and ELISA Microarray. Proteins were extracted from TSA (10, 25, 50nM) and BMP2 (10 ng/μl) treated neurosphere cultures 6, 12 or 24 h after treatment. Western Blot for pSmad1/5/8, Smad1/5/8, pStat3, Stat3, pGsk3-beta was performed. Beta-Actin was used as control ( A ). Phosphorylated proteins were quantified using an ArrayTube™ (Alere Technologies, Jena, Germany) based sandwich ELISA microarray. Concentration of pGsk3-beta ( B ) and pErk2 ( C ) in relation to total protein content is presented.

Article Snippet: For western blot analysis following primary antibodies were used: anti-pSmad1(Ser463/465)/5(Ser463/465)/8(Ser463/465) (Cell Signaling), anti-Smad1/5/8 (Santa Cruz), anti-pStat3(Tyr705) (Cell Signaling), anti-Stat3 (Cell Signaling), anti-pGsk3-beta(Ser9) (Cell Signaling), anti-Mbp (aa82-87) (AbD Serotec), anti-Gfap (DAKO), anti-Plp (aa3) (kind gift of Prof. J. Trotter, Mainz, Germany), and anti-beta-Actin (Sigma-Aldrich).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Microarray, Sandwich ELISA, Concentration Assay

Journal: Cell reports

Article Title: Inhibition of PHLPP1/2 phosphatases rescues pancreatic β-cells in diabetes

doi: 10.1016/j.celrep.2021.109490

Figure Lengend Snippet:

Article Snippet: Membranes were blocked at room temperature using mixture of 2.5% milk (Cell Signaling Technology (CST), MA, USA) and 2.5% BSA for 1h and incubated overnight at 4°C with rabbit anti-cleaved caspase-3 (#9664), rabbit anti-cleaved PARP (rat specific; #9545), rabbit anti-tubulin (#2146), rabbit anti-GAPDH (#2118), rabbit anti-β-actin (#4967), rabbit anti-GFP (#2956), rabbit anti-HA (#2367), rabbit anti-p4EBP1 (#2855), rabbit anti-pS6 (#4858), rabbit anti-pS6K (#9234), rabbit anti-Raptor (#2280), rabbit anti-MST1 (#3682), rabbit anti-AKT (#9272), rabbit anti-pAKT (#4058), rabbit anti-GST (#2625), rabbit anti-pGSK3 (#9336) (all CST), rabbit anti-PHLPP1 (RRID: AB_2750897 #22789-I-AP, Proteintech, IL, USA), rabbit anti-PHLPP2 (#A300–661A; Bethyl, TX, USA), rabbit anti-pMST1(T183) (#ab79199, Abcam, UK) and rabbit anti-GLUT2 antibody (#07–1402; Chemicon, CA, USA).

Techniques: Plasmid Preparation, Recombinant, Western Blot, Transfection, In Vivo, Bicinchoninic Acid Protein Assay, Enzyme-linked Immunosorbent Assay, In Situ, Isolation, Knock-Out, Expressing, Mutagenesis, Software, Real-time Polymerase Chain Reaction