Journal: Drug Design, Development and Therapy

Article Title:

miR-370-3p Alleviates Ulcerative Colitis-Related Colorectal Cancer in Mice Through Inhibiting the Inflammatory Response and Epithelial-Mesenchymal Transition

doi: 10.2147/dddt.s238124

Figure Lengend Snippet: Figure 3 Effect of miR-370-3p on inflammatory response and TLR4 signaling pathway in AOM/DSS-induced mice. (A) The expression of TNF-α, IL-1β, and IL-6 in colonic tissues of mice from the Sham group, UC-CRC group, Ad-control group, and Ad-miR-370-3p group was detected by qRT-PCR. (B) The production of TNF-α and PGE2 in the lysate of the tumor colon tissues were detected by ELISA. (C) The expression of TLR4 and COX-2 in the tumor colon tissues from each group was evaluated by using IHC staining. Scale bar=50 μm. (D) The expression levels of TLR4, COX-2, and pEGFR in the tumor colon tissues from each group were determined by Western blot, and the relative band intensity was analyzed.The comparison among groups was performed by One-way ANOVA. Data are presented as mean ± SD. ##p<0.01 compared with the Sham group, **p<0.01 compared with the Ad-control group.

Article Snippet: After blocking with 5% skim milk (Sangon Biotech, Shanghai China) at room temperature for 1 h, the membranes were incubated with corresponding primary antibodies incuding TLR4 (1:1000, proteintech, Wuhan, China), COX-2 (1:1000, proteintech, Wuhan, China), pEGFR (phosphorylated epidermal growth factor receptor, 1:1000, Affinity, Changzhou, Jiangsu, China), β-catenin (1:5000, proteintech, Wuhan, China), p53 (1:3000, proteintech, Wuhan, China), ki67 (1:1000, Affinity, Changzhou, Jiangsu, China), E-cadherin (1:10,000, proteintech, Wuhan, China), N-cadherin (1:5000, proteintech, Wuhan, China), Vimentin (1:5000, proteintech, Wuhan, China) and GAPDH (1:10,000, proteintech, Wuhan, China) overnight at 4° C. Then the membranes were washed by TBST and incubated with appropriate secondary HRPconjugated goat anti-rabbit or goat anti-mouse antibodies (1:3000, Solarbio, Beijing, China) for 1 h at 37 ° C. The proteins were visualized by using ECL Western Blotting Substrate (Solarbio, Beijing, China).

Techniques: Expressing, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Western Blot, Comparison

Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: Translational Psychiatry

Article Title: Progranulin improves neural development via the PI3K/Akt/GSK-3β pathway in the cerebellum of a VPA-induced rat model of ASD

doi: 10.1038/s41398-022-01875-4

Figure Lengend Snippet: Representative Western blots ( A , C ) and densitometric quantification ( B , D ) of p-Akt (Ser473), Akt, p-GSK-3β (Ser9) and GSK-3β expressions in the cerebellum. Representative Western blots ( E ) and densitometric quantification ( F – H ) of M6PR, SORT1 and PSAP. Data are expressed as mean ± SEM. One-way ANOVA followed by Tukey’s post hoc ( B , D ); Student’s t test ( F , H ); Mann–Whitney test ( G ). (* P < 0.05, ** P < 0.01, *** P < 0.001, sample sizes ( n ): n = 5/group).

Article Snippet: The antibodies used were anti-PGRN (catalogue: 18410-1-AP; Proteintech, China), anti-Caspase-3 (catalogue: 19677-1-AP; Proteintech, China), anti-Bcl-2 (catalogue: ab196495; Abcam, UK), anti-Bax (catalogue: 50599-2-Ig; Proteintech, China), anti-postsynaptic density protein 95 (PSD95) (catalogue: 20665-1AP; Proteintech, China), anti-synaptophysin (SYP) (catalogue: 17785-1-AP; Proteintech, China), anti-phosphorylated-Akt (ser473) (catalogue: AF0016; Affinity Biosciences, USA), anti-Akt (catalogue: 10176-2-AP; Proteintech, China), anti-phosphorylated-GSK-3β (ser 9) (catalogue: bs-2066R; Bioss Biotechnology, China), anti-GSK-3β (catalogue: bsm-33293M; Bioss Biotechnology, China), anti-mannose 6-phosphate receptor (M6PR) (catalogue: ab124767; Abcam, UK), anti-sortilin (SORT1) (catalogue: ab16640; Abcam, UK), anti-prosaposin (PSAP) (catalogue: A1819; ABclonal, China), anti-GADPH (catalogue: AF7021; Affinity Biosciences, USA) and anti-β-actin (catalogue: 20536-1-AP; Proteintech, China).

Techniques: Western Blot, MANN-WHITNEY

Journal: The FASEB Journal

Article Title: Mitophagy protects against statin‐mediated skeletal muscle toxicity

doi: 10.1096/fj.201900807rr

Figure Lengend Snippet: Figure 1. Simvastatin blunts Akt-mTOR signaling and induces autophagy in C2C12 myotubes. A) Western blot analysis and quantification of key activation markers of the Akt-mTOR pathway [phosphorylated Akt (pAkt) Ser473 and phosphorylated ribosomal S6 (pS6) Ser235 and Ser236] and LC3 after 24 h simvastatin (2 mM) (n = 3/group). B, C) Effects on Akt-mTOR pathway and LC3 levels by coadministration of simvastatin (2 mM) with 100 mM mevalonate for 24 h (n = 4/group) (B); phase contrast images of cells; original magnification, 320 (C). Black bars indicate DMSO control; gray bars indicate simvastatin treatment. AU, arbitrary units; Mev, mevalonate. *P , 0.05.

Article Snippet: The membranes were blocked with 5% nonfat dry milk for 1 h, then incubated with 1:500–1:1000 diluted primary antibodies against microtubuleassociated proteins 1A and 1B light chain 3B (LC3), phosphorylated protein kinase B (Akt) Ser473, Akt, phosphorylated S6 Ser235 and Ser236, S6, cleaved caspase-3, FoxO3a, cytochrome c oxidase subunit 4 (COX-IV; Cell Signaling Technology, Danvers, MA, USA), p62/sequestosome-1 (SQSTM1; Proteintech, Rosemont, IL, USA), Rho, and Lamin B1 (Santa Cruz Biotechnology, Dallas, TX, USA) at 4°C shaking overnight.

Techniques: Western Blot, Activation Assay, Control

Journal: Oncology reports

Article Title: Introduction of exogenous wild‑type p53 mediates the regulation of oncoprotein 18/stathmin signaling via nuclear factor‑κB in non‑small cell lung cancer NCI‑H1299 cells.

doi: 10.3892/or.2019.6964

Figure Lengend Snippet: Figure 1. Introduction of exogenous p53wt decreases Op18/stathmin expres- sion and phosphorylation of NCI‑H1299 cells. (A) Successfully expressed p53 downregulates the levels of Op18/stathmin in NCI‑H1299 cells. (B) p53 inhibits Op18/stathmin phosphorylation at the Ser25 and Ser63 sites. Op18, oncoprotein 18; p53wt, wild‑type p53; C3, pEGFP‑C3.

Article Snippet: The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (which was diluted at 1:3,000 for the detection of stathmin, NF-κB, IL-6 and IL-10, and at 1:2,500 for detecting caspase-9, Bcl-2, p-stathmin-Ser25, -Ser63, phospho-Thr161-CDC2, CDC2 and ERK; cat. no. sc-2004) and rabbit anti-mouse IgG (which was diluted at 1:3,000 for the detection of β-actin and p53, and at 1:2,500 for analyzing caspases-3, -8, Bcl-2-associated X and phosphor-ERK; cat. no. sc-358914) (Santa Cruz Biotechnology, Inc.).

Techniques: Phospho-proteomics

Journal: Oncology reports

Article Title: Introduction of exogenous wild‑type p53 mediates the regulation of oncoprotein 18/stathmin signaling via nuclear factor‑κB in non‑small cell lung cancer NCI‑H1299 cells.

doi: 10.3892/or.2019.6964

Figure Lengend Snippet: Figure 2. Exogenous p53wt introduction attenuates the capacities of proliferation and colony formation in NCI‑H1299 cells. (A) Cell proliferation curves of the three groups at different time points. (B) Histograms demonstrate that p53wt introduction significantly inhibits cell proliferation, compared with the two control groups, at the three time points. (C) Images depict the status of colony formation in the three groups. (D) Histograms demonstrate the differences in the colony formation ratios of the three groups. Original magnification, x10. **P<0.01. p53wt, wild‑type p53; C3, pEGFP‑C3.

Article Snippet: The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (which was diluted at 1:3,000 for the detection of stathmin, NF-κB, IL-6 and IL-10, and at 1:2,500 for detecting caspase-9, Bcl-2, p-stathmin-Ser25, -Ser63, phospho-Thr161-CDC2, CDC2 and ERK; cat. no. sc-2004) and rabbit anti-mouse IgG (which was diluted at 1:3,000 for the detection of β-actin and p53, and at 1:2,500 for analyzing caspases-3, -8, Bcl-2-associated X and phosphor-ERK; cat. no. sc-358914) (Santa Cruz Biotechnology, Inc.).

Techniques: Control

Journal: Oncology reports

Article Title: Introduction of exogenous wild‑type p53 mediates the regulation of oncoprotein 18/stathmin signaling via nuclear factor‑κB in non‑small cell lung cancer NCI‑H1299 cells.

doi: 10.3892/or.2019.6964

Figure Lengend Snippet: Figure 3. p53 inhibits migration of NCI‑H1299 cells. (A) Images depict the status of cell migration in the 2‑dimensional plane (original magnification, x4). (B) Transwell analysis demonstrates cell migration in the 3‑dimensional space (original magnification, x40). (C) Histograms depict the differences in the number of migrated cells in the three groups. **P<0.01. p53wt, wild‑type p53; C3, pEGFP‑C3.

Article Snippet: The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (which was diluted at 1:3,000 for the detection of stathmin, NF-κB, IL-6 and IL-10, and at 1:2,500 for detecting caspase-9, Bcl-2, p-stathmin-Ser25, -Ser63, phospho-Thr161-CDC2, CDC2 and ERK; cat. no. sc-2004) and rabbit anti-mouse IgG (which was diluted at 1:3,000 for the detection of β-actin and p53, and at 1:2,500 for analyzing caspases-3, -8, Bcl-2-associated X and phosphor-ERK; cat. no. sc-358914) (Santa Cruz Biotechnology, Inc.).

Techniques: Migration

Journal: Oncology reports

Article Title: Introduction of exogenous wild‑type p53 mediates the regulation of oncoprotein 18/stathmin signaling via nuclear factor‑κB in non‑small cell lung cancer NCI‑H1299 cells.

doi: 10.3892/or.2019.6964

Figure Lengend Snippet: Figure 4. Western blot analysis of the effects of exogenous p53 on the activi- ties of ERK and CDC2 kinases and transcription factor NF‑κB in NCI‑H1299 cells. (A) The expression and phosphorylation of ERK and CDC2 following p53 induction. (B) p53 inhibits the expression of NF‑κB. (C) Concentration gradients of NF‑κB inhibitor PDTC treatment. p53wt, wild‑type p53; C3, pEGFP‑C3; CDC2, cyclin‑dependent 2; ERK, extracellular signal‑regulated kinase; NF‑κB, nuclear factor‑κB; PDTC, pyrrolidine dithiocarbamate; p, phospho.

Article Snippet: The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (which was diluted at 1:3,000 for the detection of stathmin, NF-κB, IL-6 and IL-10, and at 1:2,500 for detecting caspase-9, Bcl-2, p-stathmin-Ser25, -Ser63, phospho-Thr161-CDC2, CDC2 and ERK; cat. no. sc-2004) and rabbit anti-mouse IgG (which was diluted at 1:3,000 for the detection of β-actin and p53, and at 1:2,500 for analyzing caspases-3, -8, Bcl-2-associated X and phosphor-ERK; cat. no. sc-358914) (Santa Cruz Biotechnology, Inc.).

Techniques: Western Blot, Expressing, Phospho-proteomics, Concentration Assay

Journal: Oncology reports

Article Title: Introduction of exogenous wild‑type p53 mediates the regulation of oncoprotein 18/stathmin signaling via nuclear factor‑κB in non‑small cell lung cancer NCI‑H1299 cells.

doi: 10.3892/or.2019.6964

Figure Lengend Snippet: Figure 5. p53 inhibits autocrine IL‑10 and IL‑10 expression, but does not exert any effects on IL‑6 in NCI‑H1299 cells. (A) ELISA analysis demonstrates the difference in the levels of autocrine IL‑10 among the three groups. (B) Autocrine IL‑6 ELISA assays. (C) Western blot analysis depicts the expression of IL‑10 following p53 induction. (D) Blocking of nuclear factor‑κB with PDTC attenuated IL‑10 expression in a concentration‑dependent manner. **P<0.01. p53wt, wild‑type p53; C3, pEGFP‑C3; IL, interleukin; PDTC, pyrrolidine dithiocarbamate.

Article Snippet: The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (which was diluted at 1:3,000 for the detection of stathmin, NF-κB, IL-6 and IL-10, and at 1:2,500 for detecting caspase-9, Bcl-2, p-stathmin-Ser25, -Ser63, phospho-Thr161-CDC2, CDC2 and ERK; cat. no. sc-2004) and rabbit anti-mouse IgG (which was diluted at 1:3,000 for the detection of β-actin and p53, and at 1:2,500 for analyzing caspases-3, -8, Bcl-2-associated X and phosphor-ERK; cat. no. sc-358914) (Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Blocking Assay

Journal: Oncology reports

Article Title: Introduction of exogenous wild‑type p53 mediates the regulation of oncoprotein 18/stathmin signaling via nuclear factor‑κB in non‑small cell lung cancer NCI‑H1299 cells.

doi: 10.3892/or.2019.6964

Figure Lengend Snippet: Figure 6. Analysis of growth and apoptosis in NCI‑H1299 cells following p53 introduction. (A) The upper images depict the status of cell growth and the lower panels depict the cell cycle distribution and apoptosis ratios of the three groups as determined by fluorescence‑activated cell sorting assays. (B) p53 initiates the activation of caspase‑3 and ‑9. (C) p53 decreases Bcl‑2 expression levels and increases Bax expression levels. Original magnification, x10. p53wt, wild‑type p53; C3, pEGFP‑C3; Bcl‑2, B‑cell lymphoma 2; Bax, Bcl‑2‑associated X.

Article Snippet: The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (which was diluted at 1:3,000 for the detection of stathmin, NF-κB, IL-6 and IL-10, and at 1:2,500 for detecting caspase-9, Bcl-2, p-stathmin-Ser25, -Ser63, phospho-Thr161-CDC2, CDC2 and ERK; cat. no. sc-2004) and rabbit anti-mouse IgG (which was diluted at 1:3,000 for the detection of β-actin and p53, and at 1:2,500 for analyzing caspases-3, -8, Bcl-2-associated X and phosphor-ERK; cat. no. sc-358914) (Santa Cruz Biotechnology, Inc.).

Techniques: FACS, Activation Assay, Expressing

Journal: Oncology reports

Article Title: Introduction of exogenous wild‑type p53 mediates the regulation of oncoprotein 18/stathmin signaling via nuclear factor‑κB in non‑small cell lung cancer NCI‑H1299 cells.

doi: 10.3892/or.2019.6964

Figure Lengend Snippet: Figure 7. p53 induction promotes the sensitivity of NCI‑H1299 cells to Taxol. The upper panel depicts the cell growth, and the lower panel depicts the FACS analysis of Taxol treatment. Original magnification, x10. p53wt, wild‑type p53; C3, pEGFP‑C3.

Article Snippet: The secondary antibodies used were horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (which was diluted at 1:3,000 for the detection of stathmin, NF-κB, IL-6 and IL-10, and at 1:2,500 for detecting caspase-9, Bcl-2, p-stathmin-Ser25, -Ser63, phospho-Thr161-CDC2, CDC2 and ERK; cat. no. sc-2004) and rabbit anti-mouse IgG (which was diluted at 1:3,000 for the detection of β-actin and p53, and at 1:2,500 for analyzing caspases-3, -8, Bcl-2-associated X and phosphor-ERK; cat. no. sc-358914) (Santa Cruz Biotechnology, Inc.).

Techniques: