Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Qingjie Fuzheng Granule Inhibits EMT and Induces Autophagy in Colorectal Cancer via mTOR Signaling Pathways

doi: 10.1155/2021/9950499

Figure Lengend Snippet: The protein expression of PI3K, p-PI3K, AKT, p-AKT, AMPK, p-AMKP, mTOR, and p-mTOR in tumor cells of CRC xenograft mice was detected by western blot in each group ( X ¯ ± S , n = 4). β -Actin was used as the internal control. ∗ P < 0.05, QFG treatment group vs. control group.

Article Snippet: 5% nonfat dry milk was used to block the PVDF membranes for 1 h, and then primary antibodies beclin-1 (cat. no. 11306-1-AP), p62 (cat. no. 18420-1-AP), E-cadherin (cat. no. 20874-1-AP), N-cadherin (cat. no. 22018-1-AP), vimentin (cat. no. 60330-1-AP), TWIST1 (cat. no. 25465-1-AP), PI3K (cat. no. 13329-1-AP), AKT (cat. no. 10176-2-AP), AMPK (cat. no. 10929-2-AP), mTOR (cat. no. 20657-1-AP) (1 : 1000, Proteintech, USA), p-PI3K (cat. no. ab-110021) and p-AKT (cat. no. ab-15285) (1 : 1000, Abcam, CA, USA), LC3-II (cat. no. #3868), p-AMPK (cat. no. #50081), p-mTOR (cat. no. #5536), and β -actin (cat. no. #4967) (1 : 1000, Cell Signaling Technology) were added at 4°C overnight.

Techniques: Expressing, Western Blot

Journal: Clinical and Translational Medicine

Article Title: Uterus globulin associated protein 1 (UGRP1) binds podoplanin (PDPN) to promote a novel inflammation pathway during Streptococcus pneumoniae infection

doi: 10.1002/ctm2.850

Figure Lengend Snippet: UGRP1 enhances expression of TLR2, NOD2, MyD88 and NLRP3 by activation of NF‐κB. (A) PEMs were treated with the protein synthesis inhibitor CHX (10 μM) for 1 h followed by Pam3CSK4 stimulation with BSA or UGRP1 (0.5 μg/ml) for 6 h to measure Il1b mRNA by qRT‐PCR ( n = 3). (B) The relative mRNA levels of gene from TLRs and NLRs pathway in PEMs after UGRP1 (0.5 μg/ml) treatment for 3 h ( n = 3). (C) PEMs were treated with UGRP1 (0.5 μg/ml) for 3 or 6 h to check Tlr2, MyD88, Nod2 and Nlrp3 mRNA by qRT‐PCR ( n = 3). (D) PEMs were treated with UGRP1 (0.5 μg/ml) for the indicated periods to check NLRP3, TLR2, MyD88 and NOD2 protein levels by immunoblot analysis. (E) PEMs were treated with the NF‐κB inhibitor PDTC (100 μM) for 1 h followed by BSA or UGRP1 (0.5 μg/ml) treatment for 6 h to measure Tlr2 and Nod2 mRNA by qRT‐PCR ( n = 3). (F) PEMs were treated with UGRP1 (0.5 μg/ml) for 1 h and the relative amount of Tlr2 and Nod2 DNA binding to p65 was analysed by ChIP assay. (G) MyD88 KO PEMs were treated with UGRP1 (0.5 μg/ml) for 6 h to check Tlr2 and Nod2 mRNA by qRT‐PCR ( n = 3). (H) PEMs were treated with IRAK1/4 inhibitor IRAK‐1‐4 Inhibitor I (10 μM), TAK1 inhibitor Takinib (10 μM), IKKβ inhibitor LY2409881 trihydrochloride (5 μM) or p‐IκBα inhibitor BAY 11–7082 (20 μM) for 1 h followed by UGRP1 (0.5 μg/ml) treatment for 6 h to measure Tlr2 and Nod2 mRNA by qRT‐PCR ( n = 3). * p < .05, ** p < .01 and *** p < .001, using a two‐tailed, unpaired Student's t ‐test (G), one‐way analysis of variance (ANOVA) with Holm–Sidak's multiple comparisons test (C and H) or two‐way ANOVA with Holm–Sidak's multiple comparisons test (A and E). Data from at least three independent experiments (mean ± SD) or representative data (D). Please also see Figure

Article Snippet: IRAK‐1‐4 Inhibitor I , MCE , Cat# HY‐13329.

Techniques: Expressing, Activation Assay, Quantitative RT-PCR, Western Blot, Binding Assay, Two Tailed Test

Journal: Clinical and Translational Medicine

Article Title: Uterus globulin associated protein 1 (UGRP1) binds podoplanin (PDPN) to promote a novel inflammation pathway during Streptococcus pneumoniae infection

doi: 10.1002/ctm2.850

Figure Lengend Snippet:

Article Snippet: IRAK‐1‐4 Inhibitor I , MCE , Cat# HY‐13329.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Cell Counting, Software, Real-time Polymerase Chain Reaction