Journal: Frontiers in Pharmacology

Article Title: Periplanta americana extract regulates the Th17/Treg cell balance via Notch1 in ulcerative colitis

doi: 10.3389/fphar.2024.1534772

Figure Lengend Snippet: PAE regulates the balance of Th17/Treg cells in UC mice. (A) Flow cytometry analysis of Th17 (CD4 + IL-17A + ) and Treg (CD4 + CD25 + Foxp3 + ) cells in spleen. (B) Western blot images of RORγt and Foxp3 in the colon. (C, D) Analysis of gray-scale of RORγt and Foxp3 protein expression. (E) Levels of TGF-β in peripheral blood. (F) Immunofluorescence images of RORγt and IL-17A in colon (×100). (n = 4–6, * P < 0.05 and ** P < 0.01 versus control group, # P < 0.05 and ## P < 0.01 versus model group).

Article Snippet: After blocking with 5% skim milk for 1.5 h (h) at room temperature, the membranes were incubated overnight at 4°C with different primary antibodies against Occludin (13409-1-AP, Proteintech), Claudin 1 (13050-1-A, Proteintech), ZO1 (TA5145F, Abmart; 21773-1-A, Proteintech), Foxp3 (Ab215206, Abcam), RORγt (13205-1-AP, Proteintech), Math1 (21215-1-A, Proteintech), Notch1 (T55244S, Abmart; 20687-1-AP, Proteintech), Hes1(11988S, CST) and β-actin (ab179467; Abcam).

Techniques: Flow Cytometry, Western Blot, Expressing, Immunofluorescence, Control

Journal: Frontiers in Pharmacology

Article Title: Periplanta americana extract regulates the Th17/Treg cell balance via Notch1 in ulcerative colitis

doi: 10.3389/fphar.2024.1534772

Figure Lengend Snippet: PAE attenuates the disruption of tight junction proteins in HCoEpic cells by regulating activated Notch1-mediated Th17/Treg imbalance in vitro . (A) Effect of VPA on the viability of Jurkat T cells analyzed with CCK8 assay. (B) Effect of PAE on the viability of Jurkat cells analyzed with CCK-8 assay. (C) Western blot images of Notch1, Math1, RORγt, and Foxp3 in Jurkat T cells. (D–G) Analysis of gray-scale of Notch1, Math1, RORγt, and Foxp3 protein expression. (H) After notch1-activated Jurkat T cells were co-cultured with HCoEpic cells, Western blot images of occluding, claudin1, and ZO1 in HCoEpic cells. (I–K) Analysis of gray-scale of occluding, claudin1, and ZO1 protein expression. (n = 5–8, * P < 0.05 and ** P < 0.01 versus control group, # P < 0.05 and ## P < 0.01 versus VPA group).

Article Snippet: After blocking with 5% skim milk for 1.5 h (h) at room temperature, the membranes were incubated overnight at 4°C with different primary antibodies against Occludin (13409-1-AP, Proteintech), Claudin 1 (13050-1-A, Proteintech), ZO1 (TA5145F, Abmart; 21773-1-A, Proteintech), Foxp3 (Ab215206, Abcam), RORγt (13205-1-AP, Proteintech), Math1 (21215-1-A, Proteintech), Notch1 (T55244S, Abmart; 20687-1-AP, Proteintech), Hes1(11988S, CST) and β-actin (ab179467; Abcam).

Techniques: Disruption, In Vitro, CCK-8 Assay, Western Blot, Expressing, Cell Culture, Control

Journal: Cell reports

Article Title: RORγt phosphorylation protects against T cell-mediated inflammation

doi: 10.1016/j.celrep.2022.110520

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: RORg pS203 / RORgt pS182 , Rockland-Inc , Cat# 600-401-GR8.

Techniques: Functional Assay, Recombinant, Cell Isolation, Retroviral, Software, Sequencing, Cell Culture

Journal: Kidney international

Article Title: Retinoic acid receptor responder1 promotes development of glomerular diseases via the Nuclear Factor-κB signaling pathway.

doi: 10.1016/j.kint.2021.05.036

Figure Lengend Snippet: Figure 2 | Rarres1 overexpression activates receptor tyrosine kinase Axl and consequently enhances nuclear factor kB (NF-kB) and transforming growth factor (TGF)–b1 signaling. (a) Relative mRNA levels of human Rarres1 in stable Rarres1 overexpressing (Rarres1) and Rarres1 short interfering RNA (siRNA) transfected (siRarres1) cultured human podocytes. Data are expressed as a fold change of Rarres1 expression in the transfected cells compared with the endogenous Rarres1 mRNA level. ###P < 0.001 versus control (ctrl.) or si-negative control (scramble). (b) Relative mRNA levels of epithelial-mesenchymal transition (EMT)–related genes in podocytes overexpressing Rarres1 or transfected with Rarres1 siRNA (siRarres1). ##P < 0.01, ###P < 0.001 versus control (ctrl.) or ***P < 0.001 versus si-negative control (scramble) (Rarres1, n ¼ 6; siRarres1, n ¼ 4). (c) Representative Western blot documents and summarized data showing the relative protein levels of pP65 in isolated nuclear fractions of podocytes treated with 10 ng/ml TGF-b1. **P < 0.01, ***P < 0.001 versus control podocytes (ctrl.) (n ¼ 3). (d) Representative Western blot documents and summarized data showing the relative protein levels of pP65 in isolated nuclear fractions of podocytes treated with 10 ng/ml TGF-b1 3 mmol/l R428, n ¼ 2. (e) Representative Western blot showing that overexpression of Rarres1 activates Axl constitutively. *P < 0.05 (n ¼ 2). (f) Relative mRNA levels of EMT-related genes in podocytes overexpressing Rarres1 (Rarres1) or control podocytes (ctrl.) transfected with siRNA for Axl (siAxl), si-negative control (scramble), ##P < 0.01, ###P < 0.001 versus scramble, n ¼ 3, or treated with 3 mmol/l R428. ##P < 0.01, ###P < 0.001 versus ctrl., **P < 0.01, ***P < 0.001 Q39 versus Rarres1, n ¼ 4. Data are expressed as means SD. Student’s t test was employed for comparisons between 2 groups. UT, untreated.

Article Snippet: Transfection of short interfering RNA (siRNA) for Axl/Rarres1 was performed using siRNA from Origene.

Techniques: Over Expression, Small Interfering RNA, Transfection, Cell Culture, Expressing, Control, Negative Control, Western Blot, Isolation