phospho yb 1  (Cell Signaling Technology Inc)

Journal: Molecular Oncology

Article Title: The subcellular distribution of phosphorylated Y‐box‐binding protein‐1 at S102 in colorectal cancer patients, stratified by KRAS mutational status and clinicopathological features

doi: 10.1002/1878-0261.70060

Figure Lengend Snippet: Demonstration of S102 YB‐1 immunohistochemistry staining intensity in the semiquantitative system in (A) cytoplasm (0 ( n = 1), (+) ( n = 14), 1+ ( n = 11), 2+ ( n = 10)) and (B) nucleus (− ( n = 19), + ( n = 17)). Scale bar 25 μm. Original magnification ×400.

Article Snippet: Immunohistochemistry (IHC) was performed with rabbit anti‐P‐YB‐1 S102 (Cell Signaling, #2900) on an automated immunostainer (Leica Bond‐MAX; Leica Biosystems, Wetzlar, Germany) according to the company's protocols.

Techniques: Immunohistochemistry, Staining

Journal: Molecular Oncology

Article Title: The subcellular distribution of phosphorylated Y‐box‐binding protein‐1 at S102 in colorectal cancer patients, stratified by KRAS mutational status and clinicopathological features

doi: 10.1002/1878-0261.70060

Figure Lengend Snippet: Subcellular staining of YB‐1 phosphorylation in CRC patient tissues ( n = 36). (A, B) Immunohistochemistry and immunofluorescence staining. Original magnification ×50 (top left images). (C) Negative staining of phospho‐YB‐1 in the cytoplasm and in the nucleus ( n = 1). (D) Positive staining of S012 YB‐1 (P‐YB‐1) in the cytoplasm but not in the nucleus ( n = 18). (E) Positive staining of P‐YB1 both in the cytoplasm and nucleus ( n = 17). Blue = AF647 (total YB‐1), Red = Cy3 (S102 P‐YB1), Green = YoPro1 (nucleus).

Article Snippet: Immunohistochemistry (IHC) was performed with rabbit anti‐P‐YB‐1 S102 (Cell Signaling, #2900) on an automated immunostainer (Leica Bond‐MAX; Leica Biosystems, Wetzlar, Germany) according to the company's protocols.

Techniques: Staining, Phospho-proteomics, Immunohistochemistry, Immunofluorescence, Negative Staining

Journal: Molecular Oncology

Article Title: The subcellular distribution of phosphorylated Y‐box‐binding protein‐1 at S102 in colorectal cancer patients, stratified by KRAS mutational status and clinicopathological features

doi: 10.1002/1878-0261.70060

Figure Lengend Snippet: The staining of YB‐1 phosphorylation in CRC patient tissues. (A) Representative IHC staining of the staining intensity of phosphorylated YB‐1 at S102 in normal mucosa vs. adenocarcinoma of the same patient. (B, C) Summarizing cross tables showing the expression of S102 YB‐1 in the cytoplasm vs. nucleus in cancerous (B) and in normal tissues (C) included in this study. (D) Relative representation of S102 YB‐1‐negative and ‐positive fractions of samples.

Article Snippet: Immunohistochemistry (IHC) was performed with rabbit anti‐P‐YB‐1 S102 (Cell Signaling, #2900) on an automated immunostainer (Leica Bond‐MAX; Leica Biosystems, Wetzlar, Germany) according to the company's protocols.

Techniques: Staining, Phospho-proteomics, Immunohistochemistry, Expressing

Journal: Molecular Oncology

Article Title: The subcellular distribution of phosphorylated Y‐box‐binding protein‐1 at S102 in colorectal cancer patients, stratified by KRAS mutational status and clinicopathological features

doi: 10.1002/1878-0261.70060

Figure Lengend Snippet: Associations between S102 YB‐1 levels and KRAS/BRAF mutations, as well as additional clinicopathological parameters, in CRC patients. (A) Distribution of CRC localization in the study population (gray area). Relative proportion of KRAS/BRAF mutation and nuclear P‐YB‐1 at indicated locations in the colon ( n = 36). (B) Bar graph showing the relative number of samples with nuclear P‐YB‐1 dependent on KRAS/BRAF mutation status ( KRAS mut n = 15, wt n = 12, BRAS mut n = 9). (C, D) Bar graph showing the absolute number of cases with and without nuclear P‐YB‐1, dependent on KRAS mutation status (C) and BRAF mutation status (D) ( n = 36). (E, F) Impact of TNM staging, microsatellite status, grading, and lymphangio/vascular invasion on the phosphorylation status of YB‐1. Bar graphs show the proportion of samples with nuclear P‐YB1 in the T2–4 stage (E), N0, N1, and N2 subgroups (E), M0 and M1 subgroups, MSS and MSI‐High subgroups, G2 and G3 subgroups (F), L0 and L1 subgroups, V0 and V1 subgroups (F) on the intensity of phosphorylated YB‐1 in the nucleus. (G) Bar graph showing the proportions of S102 YB‐1 in the nucleus and the cytoplasm in UICC stages I–IV (G) ( n = 36). mut, mutated; n.s., non‐significant; wt, wildtype. For statistical analysis (B–G), Fisher's exact test was applied.

Article Snippet: Immunohistochemistry (IHC) was performed with rabbit anti‐P‐YB‐1 S102 (Cell Signaling, #2900) on an automated immunostainer (Leica Bond‐MAX; Leica Biosystems, Wetzlar, Germany) according to the company's protocols.

Techniques: Mutagenesis, Phospho-proteomics

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Fisetin induces DNA double-strand break and interferes with the repair of radiation-induced damage to radiosensitize triple negative breast cancer cells

doi: 10.1186/s13046-022-02442-x

Figure Lengend Snippet: Effect of fisetin on phosphorylation of YB-1 and AKT is cell line dependent. The TNBC cells were treated with indicated concentrations of fisetin for 24 h. Thereafter, protein samples were extracted and loaded into a SDS-PAGE. The level of phosphorylation of YB-1 (S102) and AKT (S473) were detected by Western blotting. Blots were stripped and incubated with antibody against YB-1 and AKT1, respectively. Actin was detected from the YB-1 detected membrane without stripping as a loading control. The histograms represent the mean densitometry values ± SD of phospho-YB-1 to actin, YB-1 to actin and phospho-AKT to AKT1 normalized to 0 µM condition from 3 independent experiments. The asterisks indicate significant fisetin mediated changes on YB-1 and AKT phosphorylation (* p < 0.05, ** p < 0.01, *** p < 0.001), **** p < 0.0001; students t-test). SD: standard deviation

Article Snippet: Primary antibodies against YB-1 (#42,042), phospho-YB-1 (S102) (#2900), phospho-RSK (T359/S363) (#9344), RSK1/RSK2/RSK3 (#9355), phospho-AKT (S473) (#9271) and p62 (#8025) were purchased from Cell Signaling Technology (Frankfurt, Germany).

Techniques: Phospho-proteomics, SDS Page, Western Blot, Incubation, Membrane, Stripping Membranes, Control, Standard Deviation