Journal: Cell proliferation

Article Title: Tetrahedral Framework Nucleic Acid Relieves Sepsis-Induced Intestinal Injury by Regulating M2 Macrophages.

doi: 10.1111/cpr.13803

Figure Lengend Snippet: FIGURE 4 | Treatment of tFNAs activated the Mertk signal pathway. (A) Representative images of immunofluorescent staining for tFNAs (red) and DAPI (blue) in the colon and ileum (Scale bar = 20 μm). (B) Immunohistochemical staining of Mertk in colon (scale bar: 100 μm). (C–F) Western blotting detection of Mertk, p-Mertk, STAT1, p-STAT1, SOCS1, SOCS3 expression in the colon. (G) Quantitative analysis of the protein expression levels. (H) Schematic diagram of the inhibition of Mertk altering macrophage activation by tFNAs. (I–L) Western blotting detection of Mertk, p- Mertk, STAT1, p-STAT1, SOCS1, SOCS3 expression in the colon. (M) Quantitative analysis of the protein expression levels. Data were represented as mean ± SD, *p < 0.05, **p < 0.01.

Article Snippet: The antibody for ZO- 1 was from Wuhan Servicebio; the antibody for occludin was from Abcam; the antibodies for Mer, STAT1, p- STAT1, SOCS1 and SOCS3 were from Cell Signalling Technology; the antibody for p- Mer (Tyr749) was from Thermo Fisher Scientific; and UNC2250 was purchased from MCE.

Techniques: Staining, Immunohistochemical staining, Western Blot, Expressing, Inhibition, Activation Assay

Journal: Viruses

Article Title: ZIKV Strains Elicit Different Inflammatory and Anti-Viral Responses in Microglia Cells

doi: 10.3390/v15061250

Figure Lengend Snippet: Infection of BV2 microglia cells with ZIKV PE243 and ZIKV MR766 strains differentially modulate the pro-inflammatory miRNA-155 and anti-inflammatory and anti-viral factors. BV2 cells were infected with both ZIKV PE243 or ZIKV MR766 , with an MOI of 1, at 12, 24, 48, and 72 hpi using mock-treated (Mock) as a negative control and LPS + ATP (LPS) as a positive control. ( a ) miRNA-155, ( b ) SOCS1, ( d ) SOCS3 and ( e ) SHIP1 mRNA expression levels were measured by RT-qPCR. ∆∆Ct plotted results were normalized to hprt1 and mock values for mRNAs, and U6 and mock for miRNAs. ( c ) The protein SOCS1 expressions were analyzed by Western analysis. The membrane was stained with anti-SOCS1 and anti-β-actin as a loading control. Statistical analysis was performed by One-Way ANOVA, considering * p < 0.05, ** p < 0.01, and *** p < 0.001 as significant differences.

Article Snippet: Primary antibodies against PPAR-γ (Thermo Fisher Scientific Inc., PA3-821A) and SOCS1 (Boster Biological Technology Co. Ltd, Pleasanton, CA, USA, PA1074) were added to the membranes, and then secondary antibodies against mouse (Cell Signaling Technology, 7076S, Danvers, MA, USA) and rabbit (KPL, 04-15-06) were used.

Techniques: Infection, Negative Control, Positive Control, Expressing, Quantitative RT-PCR, Western Blot, Membrane, Staining, Control

Journal: Viruses

Article Title: ZIKV Strains Elicit Different Inflammatory and Anti-Viral Responses in Microglia Cells

doi: 10.3390/v15061250

Figure Lengend Snippet: Comparison of inflammatory, anti-inflammatory, and anti-viral marker expressions by BV2 microglial cell line after infection with Brazilian and African ZIKV strains (ZIKV PE243 and ZIKV MR766 ).

Article Snippet: Primary antibodies against PPAR-γ (Thermo Fisher Scientific Inc., PA3-821A) and SOCS1 (Boster Biological Technology Co. Ltd, Pleasanton, CA, USA, PA1074) were added to the membranes, and then secondary antibodies against mouse (Cell Signaling Technology, 7076S, Danvers, MA, USA) and rabbit (KPL, 04-15-06) were used.

Techniques: Comparison, Marker, Infection

Journal: JCI insight

Article Title: The E3 ubiquitin ligase Siah1 regulates adrenal gland organization and aldosterone secretion.

doi: 10.1172/jci.insight.97128

Figure Lengend Snippet: Figure 5. Downregulation of LXR/RXR and FXR/RXR expression and cholesterol biosynthesis correlates with induction of PIAS1 SUMO E3 ligase pro- tein in the adrenal glands of Siah1a–/– mice. (A) Heatmap of RNA-seq data from the adrenal glands of female and male WT and Siah1a KO mice, showing the top canonical pathways upregulated and downregulated in Siah1a KO mice. Selection criteria were P ≤ 0.05 and ≥2-fold change in expression. (B) PIAS1 staining (red) in sections of adrenal glands from 21-day-old WT and Siah1a KO mice. Scale bar: 100 μm. Original magnification, ×3 (insets). Images shown are representative of images from 4 different mice per group. (C) Western blot analysis of PIAS1 and tubulin expression in protein lysates of pooled adrenal

Article Snippet: Antibodies to α-tubulin (Santa Cruz, catalog sc-8035), β-actin (Santa Cruz, catalog sc-47778), PIAS1 (Cell Signaling for Western blot analysis, catalog 3550; Boster Biological Technology for mouse immunofluorescence, catalog PA2252), Flag (Millipore, catalog F1804), myc (Cell Signaling, catalog 227G), DAB2 (Cell Signaling, catalog 12906), β-catenin (Abcam, catalog ab6302), MECA 32 (BD biosciences, catalog BDB550563), eNACα (Bioss antibodies, catalog bs-2957R), SGK1 (Boster biological Technology, catalog PA2184), and V5 (Invitrogen, catalog R96025) were used according to the manufacturers’ recommendations.

Techniques: Expressing, RNA Sequencing, Selection, Staining, Western Blot

Journal: JCI insight

Article Title: The E3 ubiquitin ligase Siah1 regulates adrenal gland organization and aldosterone secretion.

doi: 10.1172/jci.insight.97128

Figure Lengend Snippet: Figure 6. Reduced activity of Siah1 I251L and increased PIAS1 expression in a patient with primary aldosteronism. (A) Cartoon representation of Siah1 surrounding the I151L mutation site (yellow sticks, blue label) that lies on a “helix-turn-helix” element that connects the dimer interface to the linker between the 2 Zn fingers (indicated by dashed black double-headed arrow). Ile151 is partly buried but also forms part of a prominent pocket (“D”; brown dashed circle). Loss-of-function mutation sites are colored red, and a red circle emphasizes how they cluster across the SBD dimer interface (part of second SBD is in gray). Three crystal forms are overlaid (see Methods and Results) to demonstrate conformational hetero- geneity in the second Zn finger (ZnF-2). For clarity, ZnF-2 is represented by a single helix (each structure is labeled by its PDB code, see Methods), but the motion is simple rigid-body rotation about the axis labeled “R,” circled in orange. The RING domain is N-terminal to ZnF-2, and its structure has not been determined. (B) Western blot analysis of PIAS1 expression in 293T cells transfected with PIAS1-V5 alone or in combination with either Siah1-myc or Siah1-I251L-myc. A cycloheximide (CHX) chase was performed for the indicated times. Blots were probed with antibodies against V5 or myc. Tubulin served as a loading control. Dashed lines denote the different degradation rate of PIAS1 coexpressed with WT Siah1 versus PIAS1 coexpressed with Siah1-I251L. The graph shows quantification of PIAS1 degradation from 5 independent experiments. *P < 0.05, compared with

Article Snippet: Antibodies to α-tubulin (Santa Cruz, catalog sc-8035), β-actin (Santa Cruz, catalog sc-47778), PIAS1 (Cell Signaling for Western blot analysis, catalog 3550; Boster Biological Technology for mouse immunofluorescence, catalog PA2252), Flag (Millipore, catalog F1804), myc (Cell Signaling, catalog 227G), DAB2 (Cell Signaling, catalog 12906), β-catenin (Abcam, catalog ab6302), MECA 32 (BD biosciences, catalog BDB550563), eNACα (Bioss antibodies, catalog bs-2957R), SGK1 (Boster biological Technology, catalog PA2184), and V5 (Invitrogen, catalog R96025) were used according to the manufacturers’ recommendations.

Techniques: Activity Assay, Expressing, Mutagenesis, Labeling, Western Blot, Transfection, Control

Journal: Pharmaceutical Biology

Article Title: Shenlian extract attenuates myocardial ischaemia-reperfusion injury via inhibiting M1 macrophage polarization by silencing miR-155

doi: 10.1080/13880209.2022.2117828

Figure Lengend Snippet: The effect of SL on SOCS3 expression. (A, B) SOCS3 gene expression and quantitative analysis by qRT-PCR. (C, D) SOCS3 proteins expression of and quantitative analysis in pMACs by western blotting. Results were presented as mean ± SD. * p < 0.05, ** p < 0.01 vs. control group, # p < 0.05, ## p < 0.01 vs. model group, n = 3.

Article Snippet: SOCS3 (Cat. No: A00274-2) antibody was purchased from BOSTER. iNOS (Cat. No: 18985-1-AP) antibody was obtained from Proteintech.

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Western Blot, Control