Journal: Endocrine, Metabolic & Immune Disorders Drug Targets

Article Title: Elucidating the Mechanism of Jisheng Shenqi Pills in the Treatment of Diabetic Kidney Disease: Network Pharmacology Combined with Experimental Verification

doi: 10.2174/0118715303339433240805045749

Figure Lengend Snippet: ( a-e ) JSP treatment improved the mRNA expression of AGE and the protein expression levels of RAGE, p-AKT/AKT and p-PI3K/PI3K in the kidney of DKD mice. ( f-h ) JSP treatment could improve the expression levels of BAX/Bcl-2 ratio, Cleaved-Caspase-3 and Bcl2 proteins in the kidney of DKD mice. ( i-k ) JSP treatment could improve the mRNA expression of IL-1β, TNF-α and MCP-1 in DKD mice kidney. ### p < 0.001 means compared with the normal control group; ## p <0.01 means compared with the normal control group; # p <0.05 indicates comparison with the normal control group; *** p <0.001 indicates that compared with the model group; ** p <0.01 indicates that compared with the model group; * p <0.05 indicates comparison with the model group.

Article Snippet: The proteins were transferred to PVDF membranes via gel electrophoresis and then blocked with a commercial blocking solution for 15 min. Next, the membranes were incubated with primary antibodies targeting RAGE (rabbit, 1:1000, Cat# YP-Ab-13666, UpingBio), PI3K (rabbit, 1:500, Cat# ET1608-70, HUABIO), p-PI3K (rabbit, 1:1000, Cat# YP-Ab-17845, UpingBio), AKT (rabbit, 1:3000, Cat# ET1609-51, HUABIO), p-AKT (rabbit, 1:2000, Cat# ET1607-73, HUABIO), BAX (rabbit, 1:5000, Cat# ER0907, HUABIO), Bcl2 (rabbit, 1:2000, Cat# ET1702-53, HUABIO), and Cleaved-Caspase-3 (rabbit, 1:1000, Cat# YP-Ab-00003, UpingBio) for 12 h at 4°C.

Techniques: Expressing, Control, Comparison

Journal: Molecular Cancer Research

Article Title: AKT1 E17K Inhibits Cancer Cell Migration by Abrogating β-Catenin Signaling

doi: 10.1158/1541-7786.mcr-20-0623

Figure Lengend Snippet: Figure 4. AKT2, but not AKT1, increases the transcriptional activity of b-catenin on the ZEB1 promoter, and decreases E-cadherin expression. A, p53ko/E17K cells were infected with a CRISPR/Cas9 lentivirus targeting AKT1 and p53ko cells were infected with a CRISPR/Cas9 lentivirus targeting AKT2. Cells lysates were then subjected to ChIP using a b-catenin antibody. Top: Western blot of whole cell lysates from the indicated p53ko/E17K cells and p53ko cells before and after AKT1 knockdown (AKT1KD) or AKT2 knockdown (AKT2KD) as specified. Bottom: Relative fold of ChIP pull-down of the ZEB1 promoter (b-catenin fold enrichment) was quantified by real-time qPCR, normalized by both input DNA and IgG values. Error bars, mean SEM (n ¼ 3). P values were determined by one-way ANOVA followed by Tukey post hoc test. , P < 0.0001. B, Western blots of lysates from MCF-10A p53ko/E17K or MCF7 PIK3CA WT cells expressing HA-tagged Myr-AKT2 and FLAG-tagged Myr-AKT1 alone and in combination. C, Western blot of lysates from p53ko cells: Left: infected with PTEN CRISPR/Cas9 lentivirus; Right: infected with PIK3CA E545K lentivirus. Cell lysate of parental MCF-10A cells was used as an E-cadherin–positive control.

Article Snippet: PIK3CA E545K–mutant cDNA (Cat#RC400348), purchased from OriGene, was inserted into the AsiS I and Mlu I sites of plasmid Lenti-C-Myc-DDK-IRES-Neo (Cat#PS100081, OriGene), to generate PIK3CA-E545K lentiviral expression vector.

Techniques: Activity Assay, Expressing, Infection, CRISPR, Western Blot, Knockdown, Positive Control

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Silencing MEG3 protects PC12 cells from hypoxic injury by targeting miR-21.

doi: 10.1080/21691401.2020.1725533

Figure Lengend Snippet: Figure 6. Silence of MEG3 activates PI3K/AKT pathway while deactivates NF-jB pathway via targeting miR-21. PC12 cells were co-transfected with si-MEG3/si-NC and miR-21 inhibitor/NC inhibitor, and then incubated in hypoxia for 24 h. The cells without transfection in normoxic condition served as control (CTRL). The expres- sion of proteins involved in (A,B) PI3K/AKT and (C,D) NF-jB pathways was tested by western blot. n ¼ 3.p < .05 represents a comparison of marked groups.

Article Snippet: Proteins were carried out by using primary antibodies specific for CyclinD1 (orb77046), p53 (orb178524), p21 (orb38089), Bcl-2 (orb378569), Bax (orb378566), pro-caspase-3 (orb13278), cleaved-caspase-3 (orb310032), PTEN (orb39110), PI3K (orb395443), p-PI3K (orb106105), AKT (orb29949), p-AKT (orb344403), IjBa (orb13487), p-IjBa (orb338946), p65 (orb344389), p-p65 (orb106211), and b-actin (orb378579, all from Biorbyt, San Francisco, CA).

Techniques: Transfection, Incubation, Control, Western Blot, Comparison

Journal: Cell Reports Medicine

Article Title: HOXC6 drives a therapeutically targetable pancreatic cancer growth and metastasis pathway by regulating MSK1 and PPP2R2B

doi: 10.1016/j.xcrm.2023.101285

Figure Lengend Snippet:

Article Snippet: Plasmid: pBabe puro pI3K p110 CAAX , Addgene , Cat# 13339.

Techniques: Western Blot, Immunohistochemistry, Immunoprecipitation, Microarray, Recombinant, Transfection, Plasmid Preparation, Membrane, Staining, Flow Cytometry, Viability Assay, Chromatin Immunoprecipitation, Phospho-proteomics, shRNA, Sequencing, Knockdown, Binding Assay, Control, Software, Expressing, Gene Expression

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: A, B. Low expression of PTEN and PIK3C2A were correlated with high risk, poor prognosis and shorter OS time. C, D. High expression of ITPA and BCL3 indicated high risk, poor prognosis and shorter OS time. Kaplan-Meier survival curves were also constructed to reveal the relationship between predicted risk of ccRCC patients and the OS time. The results showed that patients with high risk had a significantly shorter OS time than those with low risk (A-D). Green and red lines indicated low- and high-risk groups, respectively. P <0.05 was considered to be statistically significant. Cens: Censored; Event: Death; Prog. Idx.: Prognosis Index.

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing, Construct

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: IHC analysis on tissue microarrays and Kaplan-Meier survival curves were constructed to verify the relationship between PTEN A. , PIK3C2A B. , ITPA C. and BCL3 D. expression with regard to OS and DFS. (A, B) Negative expression of PTEN and PIK3C2A were correlated with shorter OS and DFS time and worse prognosis. (C, D) Positive expression of ITPA and BCL3 were correlated with shorter OS and DFS time and worse prognosis. Green and blue lines indicated positive and negative expression groups, respectively. P <0.05 was considered to be statistically significant.

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Construct, Expressing

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: A. SurvExpress was used to analyze the association of the four-gene signature with the predicted risk, survival time and prognosis. B. The gene expression level of PTEN, PIK3C2A, ITPA and BCL3 were detected in high risk and low risk group. C. Kaplan-Meier survival curves showed that patients with predicted high risk ( n = 219) had significantly shorter OS time than those with low risk ( n = 249) ( P <0.05). D. ROC analysis was performed to compare the sensitivity and specificity of the survival prediction between our models. P <0.05 was considered to be statistically significant. Cens: Censored; Event: Death; Prog. Idx.: Prognosis Index; Sur.(M): Survival status (Month).

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Gene Expression

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: Correlation between clinicopathological features and the four proteins expression in ccRCC ( n =174)

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: Univariate and multivariate Cox proportional hazard regression analyses of the association of clinicopathological characteristics and the four genes' expression levels with OS

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing

Journal: Oncotarget

Article Title: A four-gene signature predicts survival in clear-cell renal-cell carcinoma

doi: 10.18632/oncotarget.12631

Figure Lengend Snippet: Univariate and multivariate Cox proportional hazard regression analyses of the association of clinicopathological characteristics and the four genes' expression levels with DFS

Article Snippet: Subsequently, the sections were first incubated with protein blocker for 1 h at 37°C, and then incubated with anti-PTEN antibody (Ready-to-use, ZSGB-BIO, ZA-0251, China), anti-PIK3C2A antibody (1:50, Proteintech, 22028-1-AP, China), anti-ITPA antibody (1:50, Proteintech, 16134-1-AP, China) and anti-BCL3 antibody (1:50, Proteintech, 23959-1-AP, China) at 4°C overnight.

Techniques: Expressing

Journal: Food Science and Human Wellness

Article Title: Sucrose-free hawthorn leathers formulated with fructooligosaccharides and xylooligosaccharides ameliorate high-fat diet induced inflammation, glucose and lipid metabolism in liver of mice

doi: 10.1016/j.fshw.2022.03.033

Figure Lengend Snippet: Fig. 4 Effect of hawthorn leather treatment on expression levels of inflammatory cytokines and NF-κB signaling pathway in the liver tissue of mice. mRNA expression level of (A) IL-1β, (B) Nos2, (C) Cox2, (D) TLR4, (E) MyD88, and (F) NF-κB; (G) Images of western blotting; Protein expression ratio of (H) MyD88 to GAPDH and (I) NF-κB to GAPDH. Data are expressed as mean ± SD (8 mice/group). Values of each group with different letters over the bars are significantly different (P < 0.05) by analysis of one-way ANOVA followed by Tukey’s multiple range test.

Article Snippet: Then, the membrane was incubated with specific primary antibody (1 : 1 000) against MyD88 (CST, cat. number 4283S), NF-κB (p65) (CST, cat. number 8242S), PI3K (p110β) (Proteintech, cat. number 20584-1-AP), AKT (Proteintech, cat. number 10176-2-AP), Thr308 phosphorylated AKT (p-AKT) (CST, cat. number 13038T), mTOR (Proteintech, cat. number 20657-1-AP), Ser2448 phosphorylated mTOR (p-mTOR) (CST, cat. number 5536T), PPAR-γ (CST, cat. number 2435T), FASN (CST, cat. number 3180T), and HSL (CST, cat. number 18381T) overnight at 4 °C, respectively.

Techniques: Expressing, Western Blot

Journal: Brazilian Journal of Physical Therapy

Article Title: Can transcutaneous electrical nerve stimulation improve achilles tendon healing in rats?

doi: 10.1590/bjpt-rbf.2014.0107

Figure Lengend Snippet: Comparison of the quantity of collagen types III (%) in the tendon tissue between the TENS and Simulation groups. Data expressed in mean and standard deviation.

Article Snippet: The expression of collagen I and III, observed in the immunohistochemical reaction with the polyclonal antibodies for the collagen types I and III (Affinity Purified Anti-Collagen Type I and Type III, Rabbit Anti-Human, Rockland Inc., Gilbertsville, PA, USA), was analyzed by quantifying the area corresponding to each type of collagen.

Techniques: Comparison, Standard Deviation

Journal: Brazilian Journal of Physical Therapy

Article Title: Can transcutaneous electrical nerve stimulation improve achilles tendon healing in rats?

doi: 10.1590/bjpt-rbf.2014.0107

Figure Lengend Snippet: Measures of variables related to group, assessment, and results of Analysis of Variance (ANOVA).

Article Snippet: The expression of collagen I and III, observed in the immunohistochemical reaction with the polyclonal antibodies for the collagen types I and III (Affinity Purified Anti-Collagen Type I and Type III, Rabbit Anti-Human, Rockland Inc., Gilbertsville, PA, USA), was analyzed by quantifying the area corresponding to each type of collagen.

Techniques: Standard Deviation