s6 ribosomal protein 54d2  (Thermo Fisher)

Journal: EMBO Molecular Medicine

Article Title: Somatic PIK3R1 mutations in the iSH2 domain are accessible to PI3Kα inhibition

doi: 10.1038/s44321-025-00249-9

Figure Lengend Snippet: ( A ) Representative immunostainings of P-AKT T308 and P-S6RP in skin vascular malformations from three patients and skin biopsies from healthy controls, and quantification. Scale bar 50 μm. Graphs show mean ± SEM from 3 biological replicates in each group. P -values were obtained from Student’s t-tests. ( B ) Western blot and quantification of P-AKT S473 and P-S6RP levels in HeLa cells transfected with plasmids containing either PIK3R1 WT , PIK3R1 c.1735_1740del or PIK3R1 c.1372_1373dupAAA variants, treated with either vehicle or alpelisib. In all experiments, cells were stimulated with recombinant human IGF-1 (10 ng/mL) for 30’. Graphs show mean ± SEM from 4 biological replicates in each group. P -values were obtained from two-way ANOVAs; exact p -values for P-AKT/α-tubulin are: p = 3.81 × 10 −12 when comparing PIK3R1 c.1735_1740del (DMSO) and PIK3R1 WT (DMSO), p = 1.95 × 10 −5 when comparing PIK3R1 c.1372_1373dupAAA (DMSO) and PIK3R1 WT (DMSO) and p = 1.82 × 10 −7 when comparing PIK3R1 c.1372_1373dupAAA (alpelisib 1 µM) and PIK3R1 c.1372_1373dupAAA (DMSO). ( C ) Western blot and quantification of P-AKT S473 and P-S6RP levels in HeLa cells transfected with plasmids containing either PIK3R1 WT or PIK3R1 c.1699A>G, c.1703C>T variant, treated with either vehicle or alpelisib. In all experiments, cells were stimulated with recombinant human IGF-1 (10 ng/mL) for 30’. Graphs show mean ± SEM from 3 biological replicates in each group. P -values were obtained from two-way ANOVAs; exact p -values for P-AKT/α-tubulin are: p = 8.11 × 10 −9 when comparing PIK3R1 c.1699A>G, c.1703C>T (DMSO) and PIK3R1 WT (DMSO), p = 1.12 × 10 −5 when comparing PIK3R1 c.1699A>G, c.1703C>T (alpelisib 1 µM) and PIK3R1 c.1699A>G, c.1703C>T (DMSO). .

Article Snippet: S6 Ribosomal Protein (54D2) Mouse mAb , Cell Signaling , CS2317.

Techniques: Western Blot, Transfection, Recombinant, Variant Assay

Journal: EMBO Molecular Medicine

Article Title: Somatic PIK3R1 mutations in the iSH2 domain are accessible to PI3Kα inhibition

doi: 10.1038/s44321-025-00249-9

Figure Lengend Snippet: Western blot and quantification of AKT phosphorylation on residue Ser 473 and S6RP in HeLa cells transfected with plasmids containing either GFP, PIK3R1 WT , PIK3R1 c.1699A>G , PIK3R1 c.1703C>T or PIK3R1 c.1699A>G, c.1703C>T variants. In all experiments, cells were stimulated with recombinant human IGF-1 (10 ng/mL) for 30’. Graphs show mean ± SEM from 3 biological replicates in each group. P -values were obtained from two-way ANOVAs; exact p -values for P-AKT S473 /tubulin are p = 9.35 × 10 −5 when comparing PIK3R1 c.1699A>G, c.1703C>T and PIK3R1 WT ; p = 7.30 × 10 −5 when comparing PIK3R1 c.1699A>G, c.1703C>T and PIK3R1 c.1703C>T . Exact p -values for P-AKT S473 /total AKT are p = 1.17 × 10 −5 when comparing PIK3R1 c.1699A>G, c.1703C>T and PIK3R1 WT ; p = 1.09 × 10 −5 when comparing PIK3R1 c.1699A>G, c.1703C>T and PIK3R1 c.1703C>T .

Article Snippet: S6 Ribosomal Protein (54D2) Mouse mAb , Cell Signaling , CS2317.

Techniques: Western Blot, Phospho-proteomics, Residue, Transfection, Recombinant

Journal: STAR Protocols

Article Title: Analyzing efficiency of a lentiviral shRNA knockdown system in human enteroids using western blot and flow cytometry

doi: 10.1016/j.xpro.2024.103082

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-S6 ribosomal protein (54D2) antibody dilution: 1/500 , Cell Signaling Technology , Cat#2317; RRID: AB_2238583.

Techniques: Recombinant, Membrane, Staining, Protease Inhibitor, Lysis, Extraction, Bicinchoninic Acid Protein Assay, Transduction, Positive Control, shRNA, Control, Sequencing, Software, Flow Cytometry, Sterility, Transferring, Adhesive, Aerosol, Cell Counting, Electrophoresis, Microscopy, Purification