Journal: The Journal of Nutrition, Health & Aging

Article Title: Adipokine pathways are altered in hippocampus of an experimental mouse model of Alzheimer's disease

doi: 10.1007/s12603-014-0574-5

Figure Lengend Snippet: mRNA expression profile (n=5-8) of stat1, stat3, stat5a, stat5b, K-ras, sos1, socs1, socs2 and socs3 in hippocampal extracts of 3 and 6 months-old wild-type and APP/PS1 mice. (Statistical analysis was performed with one-way ANOVA, with Tukey’s post-hoc test, where * denotes p < 0.05, and with the student’s t-test, where $ denotes p < 0.05)

Article Snippet: Socs2 , suppressor of cytokine signaling 2 , 216233 , Mm00850544_g1 , .

Techniques: Expressing

Journal: Cells

Article Title: The Role of GLP1 in Rat Steatotic and Non-Steatotic Liver Transplantation from Cardiocirculatory Death Donors.

doi: 10.3390/cells8121599

Figure Lengend Snippet: Figure 5. Effect of GLP1 on inflammatory responses in LT with DCDs. (A) Protein levels of suppressors of cytokine signaling 1 (SOCS1) (A1), SOCS2 (A2), and SOCS3 (A3) in liver. (B) MPO (B1) and MDA (B2) levels in liver, Von Willebrand Factor (vWF) in plasma (B3), and nitrate, nitrite (B4), and nitrotyrosine (B5) levels in liver. For A1–A3 and B1–B5, six transplants per group were performed for each measurement. Representative Western blots for the proteins SOCS1, SOCS2, and SOCS3 and the housekeeping protein β-actin are shown at the top, and densitometric quantification of SOCS1, SOCS2, and SOCS3 protein expression normalized to β-actin (arbitrary units) is shown at the bottom. + p < 0.05 vs. LT; # p < 0.05 vs. CD+LT.

Article Snippet: After assessing transfer, the membranes were saturated in 4 mM Tris-HCl, pH 7.6, 30 mM NaCl, and 0.1% Tween 20 (TBST) containing 5% non-fat milk and incubated overnight at 4 ◦C using antibodies against GLP1 (ab23468), cyclin A (sc-239), CD-26 (also known as DPP4; sc-52642), SOCS3 (sc-9023) and transferrin (sc-52256) (Santa Cruz Biotechnology, Dallas, TX, USA), SOCS1 (ARP42148) and SOCS2 (ARP63434) (Aviva Systems Biology, San Diego, CA, USA), cyclin D1 (SAB5500090), and β-Actin (A2228) (Sigma-Aldrich).

Techniques: Clinical Proteomics, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Caveolin-1 is Associated with Tumor Progression and Confers a Multi-Modality Resistance Phenotype in Pancreatic Cancer

doi: 10.1038/srep10867

Figure Lengend Snippet: (A) shCav-1 downregulated cells in both MIAPaCa-2 and BxPC3 cell lines showed decreased phosphoJAK2 at residues Tyr1007/1008 and phosphoSTAT3 at Tyr705 as compared to scrambled control shRNA cells. Total levels of JAK2 and STAT3 are unchanged. SOCS2 and PIAS3, both inhibitors of the JAK-STAT pathway, are upregulated in Cav-1 knockdown cells. Phosphorylation of Src at Tyr416, the activation site, is decreased while phosphorylation of Src at Tyr 527, the inhibition site, is increased. GAPDH equal loading control remains unchanged. (B) Phosphorylation of JNK at Thr183/Tyr185, p38MAPK at Thr180/Tyr182 and pERK1/2 at Thr202/Tyr204 is decreased in both cell lines with knockdown of Cav-1, consistent with global reduction in proliferative pathways. Totals remain unaffected. DUSP5 levels are increased in both cell lines concomitantly. GAPDH shows equal loading control. (C) Schematic depicting the pathways affected in signaling when Cav-1 is downregulated in pancreatic cancer cell lines.

Article Snippet: Anti- cleaved caspase-9, cleaved PARP, phospho-Akt, total Akt, phospho-BRCA1, phospho-DNAPK, total BRCA1, total DNAPK, phospho-JAK2, total JAK2, phospho-STAT3, total STAT3, PIAS3, Src (Y416), Src (Y527), total Src, SOCS2, phospho-JNK, total JNK, phospho-p38, total p38, phospho-ERK1/2, total ERK1/2, alpha tubulin, phospho-H2.AX, beta actin and GAPDH antibodies were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Control, shRNA, Knockdown, Phospho-proteomics, Activation Assay, Inhibition

Journal: Oncology Reports

Article Title: m 6 A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation

doi: 10.3892/or.2020.7665

Figure Lengend Snippet: Inhibition of cell proliferation in METTL3-KO SW480 cells. (A) METTL3-KO SW480 cells showed dysregulation of several proliferation-related genes (*P<0.05, compared with the SW480 WT cells). (B) MTS assay results validated the decreased proliferation of METTL3-KO cells (*P<0.05, compared with the SW480 WT cells). (C) METTL3-KO cells showed a reduced proliferation rate, as measured by cell counting (*P<0.05, compared with the SW480 WT cells). (D) METTL3-KO cells showed a lower colony formation rate than control cells at 7 days after seeding. (E) Tumorigenic ability of METTL3-KO SW480 cells was reduced (*P<0.05, compared with the SW480 WT cells). SOCS2, suppressor of cytokine signaling 2; METTL3, methyltransferase like 3; LGR5, leucine-rich repeat-containing G protein-coupled receptor 5; SOX2, SRY-box transcription factor 2; PCNA, proliferating cell nuclear antigen; KO, knockout; SW480 A7 and SW480 A12, METTL3-KO SW480 cells.

Article Snippet: The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).

Techniques: Inhibition, MTS Assay, Cell Counting, Knock-Out

Journal: Oncology Reports

Article Title: m 6 A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation

doi: 10.3892/or.2020.7665

Figure Lengend Snippet: SOCS2 is upregulated in colon cancer and downregulated in METTL3-KO SW480 cells. (A) TCGA database analysis revealed a lower expression level of SOCS2 in colon cancer tissues than in the paired normal tissues (*P<0.05). (B) Expression levels of SOCS2 and METTL3 were negatively correlated in colon cancer tissues. (C-E) METTL3-KO upregulated SOCS2 expression at both the (C) mRNA and (D and E) protein levels in SW480 cells (*P<0.05, compared with the SW480 WT cells). SOCS2, suppressor of cytokine signaling 2; METTL3, methyltransferase like 3; KO, knockout; SW480 A7 and SW480 A12, METTL3-KO SW480 cells.

Article Snippet: The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).

Techniques: Expressing, Knock-Out

Journal: Oncology Reports

Article Title: m 6 A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation

doi: 10.3892/or.2020.7665

Figure Lengend Snippet: Methylation status of SOCS2 mRNA in SW480 cells. (A) Comparison of the SOCS2 mRNA methylation status between SW480 WT and METTL3-KO SW480 cells via MeRIP-qPCR (*P<0.05, compared with the SW480 WT cells). (B) Dose-dependent increase in SOCS2 mRNA levels in SW480 cells in response to demethylation treatment with DAA (*P<0.05, compared with non-treated group). (C and D) SOCS2 protein expression was upregulated after demethylation treatment with DAA (*P<0.05, compared with the SW380 WT DAA-group). (E) DAA appreciably decreased the m 6 A level in SOCS2 mRNA (*P<0.05, compared with the SW380 WT DAA-group). (F) METTL3-KO increased SOCS2 mRNA stability, showing a decreased mRNA decay rate (*P<0.05, compared with the SW480 WT cells). SOCS2, suppressor of cytokine signaling 2; METTL3, methyltransferase like 3; KO, knockout; SW480 A7 and SW480 A12, METTL3-KO SW480 cells; DAA, 3-deazaadenosine.

Article Snippet: The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).

Techniques: Methylation, Expressing, Knock-Out

Journal: Oncology Reports

Article Title: m 6 A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation

doi: 10.3892/or.2020.7665

Figure Lengend Snippet: Modulation of SOCS2 expression alters the proliferation ability of SW480 cells. (A-C) siRNA (si-SOCS2) was used to successfully knock down SOCS2 expression in SW480 cells. Knockdown of SOCS2 had minor impact on METTL3 expression but significantly increased LGR5 expression in SW480 cells (*P<0.05, compared with the si-Ctrl group). (D) Knockdown of SOCS2 enhanced SW480 cell proliferation, as determined by an MTS assay (*P<0.05, compared with the si-Ctrl group). (E-G) Transfection of the pCMV6-SOCS2 plasmid upregulated SOCS2 expression and decreased LGR5 expression in SW480 cells (*P<0.05, compared with the pCMV6-Entry group). SOCS2 overexpression had minor impact on METTL3 expression (P>0.05, compared with the pCMV6-Entry group). (H) SOCS2 overexpression decreased SW480 cell proliferation, as determined by an MTS assay (*P<0.05, compared with the pCMV6-Entry group). SOCS2, suppressor of cytokine signaling 2; METTL3, methyltransferase like 3; LGR5, leucine-rich repeat-containing G protein-coupled receptor 5.

Article Snippet: The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).

Techniques: Expressing, MTS Assay, Transfection, Plasmid Preparation, Over Expression

Journal: Oncology Reports

Article Title: m 6 A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation

doi: 10.3892/or.2020.7665

Figure Lengend Snippet: METTL3 modulates LGR5 transcription through m 6 A-mediated changes in SOCS2 expression rather than through direct regulatory mechanisms. (A) TCGA database analysis revealed an induction of LGR5 transcription in colon cancer tissues when compared with normal tissues (**P<0.01, compared with the normal tissues). (B) Expression levels of SOCS2 and LGR5 were negatively correlated in colon cancer tissues. (C) METTL3 knockout (KO) did not obviously impact LGR5 mRNA methylation (as measured by MeRIP-qPCR). (D) METTL3 KO had a minor effect on the LGR5 mRNA decay rate. (E) DAA treatment did not impact LGR5 expression in SW480 cells. (F) Luciferase reporter assay results confirmed that SOCS2 negatively controls LGR5 promoter-driven luciferase activity (*P<0.05, pGL3 basic-LGR5 promoter group vs. FULL SOCS2-LGR5 promoter group). METTL3, methyltransferase like 3; LGR5, leucine-rich repeat-containing G protein-coupled receptor 5; SOCS2, suppressor of cytokine signaling 2; TCGA, The Cancer Genome Atlas; SW480 A7 and SW480 A12, METTL3-KO SW480 cells; DAA, 3-deazaadenosine.

Article Snippet: The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).

Techniques: Expressing, Knock-Out, Methylation, Luciferase, Reporter Assay, Activity Assay

Journal: Oncology Reports

Article Title: m 6 A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation

doi: 10.3892/or.2020.7665

Figure Lengend Snippet: Correlations among METTL3, SOCS2 and LGR5 expression levels in CRC. (A-C) Knockdown of METTL3 using siRNA (si-METTL3#1 and si-METTL3#2) significantly upregulated SOCS2 expression but downregulated LGR5 expression in SW480 cells (*P<0.05, compared with the si-Ctrl group). (D and E) Western blotting analysis of 24 paired tissues collected from CRC patients showed high protein levels of METTL3 and LGR5 and a relatively low protein level of SOCS2 in colon cancer (T, colon tumor samples; N, adjacent normal colon tissues; *P<0.05, compared to Normal tissues). (F) Visualization of expression profiles of METTL3, SOCS2 and LGR5 in CRC through the Oncomine database. METTL3 and LGR5 were upregulated in CRC datasets. In contrast, SOCS2 was downregulated in CRC datasets. CRC, colorectal cancer; SOCS2, suppressor of cytokine signaling 2; METTL3, methyltransferase like 3; LGR5, leucine-rich repeat-containing G protein-coupled receptor 5.

Article Snippet: The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).

Techniques: Expressing, Western Blot

Journal: Oncology Reports

Article Title: m 6 A methyltransferase METTL3 maintains colon cancer tumorigenicity by suppressing SOCS2 to promote cell proliferation

doi: 10.3892/or.2020.7665

Figure Lengend Snippet: Association between patient characteristics and gene expression in 24 paired CRC cases.

Article Snippet: The pCMV6 plasmid containing the full-length SOCS2 sequence (FULL-SOCS2) was purchased from Origene (RC203163).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Favorable prognostic value of SOCS2 and IGF-I in breast cancer

doi: 10.1186/1471-2407-7-136

Figure Lengend Snippet: Kaplan-Meier curve assessment of risk of death in a cohort of 89 patients with invasive breast cancer. Overall survival is shown for patients with high and low SOCS2 and IGF-I expression, respectively. The median was taken as a cut-off. P values were determined using the log-rank test.

Article Snippet: To test for antibody specificity, lysates from COS7 cells transiently transfected with a myc-tagged SOCS2 plasmid were probed with an anti-c-myc antibody (9E10, Santa Cruz Biotechnology, Santa Cruz) and anti-SOCS2 antibodies.

Techniques: Expressing

Journal: BMC Cancer

Article Title: Favorable prognostic value of SOCS2 and IGF-I in breast cancer

doi: 10.1186/1471-2407-7-136

Figure Lengend Snippet: Comparison of SOCS2 and IGF-I expression. (A) Box blots with data from a set of seven tumors comparing the relative mRNA expression levels in tumor (CA) and normal adjacent tissue (TA). (B) SOCS2 protein and corresponding SOCS2 mRNA expression in normal and cancerous tissue of two tumor specimens. SOCS2 protein expression was assessed by immuno-blot with an antibody directed against SOCS2. Actin was used as a loading control.

Article Snippet: To test for antibody specificity, lysates from COS7 cells transiently transfected with a myc-tagged SOCS2 plasmid were probed with an anti-c-myc antibody (9E10, Santa Cruz Biotechnology, Santa Cruz) and anti-SOCS2 antibodies.

Techniques: Expressing

Journal: BMC Cancer

Article Title: Favorable prognostic value of SOCS2 and IGF-I in breast cancer

doi: 10.1186/1471-2407-7-136

Figure Lengend Snippet: Detection of SOCS2 in primary mammary carcinoma samples by immunohistochemistry. Representative immunohistochemical stainings of samples with low SOCS2 (A) and high SOCS2 (B) mRNA expression are shown. Note a strong cytoplasmatic epithelial staining for SOCS2 in tumors with high SOCS2 mRNA levels (B).

Article Snippet: To test for antibody specificity, lysates from COS7 cells transiently transfected with a myc-tagged SOCS2 plasmid were probed with an anti-c-myc antibody (9E10, Santa Cruz Biotechnology, Santa Cruz) and anti-SOCS2 antibodies.

Techniques: Immunohistochemistry, Immunohistochemical staining, Expressing, Staining