sta-21 (ochromycinone)  (Cayman Chemical)

Journal: Frontiers in Immunology

Article Title: MicroRNA-382 Promotes M2-Like Macrophage via the SIRP-α/STAT3 Signaling Pathway in Aristolochic Acid-Induced Renal Fibrosis

doi: 10.3389/fimmu.2022.864984

Figure Lengend Snippet: miR-382 indirectly activates STAT3 and gets involved in AA-induced CKD. (A) Images and quantification of Western blot for p-STAT3 Y705, p-STAT3 S727, and STAT3 in renal tissues in the time course of AAN. GAPDH served as standard. (B) Images and quantification of Western blot for p-STAT3 Y705, p-STAT3 S727, and STAT3 in renal tissues from WT and KO mice of 28 days AAN. GAPDH served as standard. (C) Immunohistochemistry staining with antibodies against p-STAT3 Y705, p-STAT3 S727, and STAT3 in renal sections between WT and KO mice of 28 days AAN. Scale bars, 200 μm. (D) Images of co-staining for F4/80, p-STAT3, and DAPI in renal sections from WT-14d and KO-14d. F4/80 was stained as green; p-STAT3 was stained as red. Nucleic was stained as blue. Scale bars, 100 μm. (E) Images of Sirius Red staining in these groups. Quantification of mean positive area for Sirius Red staining in these groups. Scale bars, 200 μm. (F) Images of Western blot for p-STAT3 Y705, p-STAT3 S727, STAT3, and α-SMA in kidney from control, AA 14 days, DMSO+ AA 14 days, and STA-21+ AA 14 days groups. GAPDH served as standard. (G, H) Quantification of western blot for STAT3 and α-SMA in control, AA 14 days, DMSO+ AA 14 days, and STA-21+ AA 14 days groups. GAPDH served as standard. (I) MFI of CD206+ macrophages in kidney from control, AA 14 days, DMSO+ AA 14 days and STA-21+ AA 14 days groups. *P < 0.05; **P < 0.01; ***P < 0.001; ANOVA.

Article Snippet: STA-21 (HY-18061, MedChemExpress, Princeton, NJ, USA) was administered intraperitoneally to animals at a dose of 0.5 mg/kg the day before AA injection (DMSO was used as control).

Techniques: Western Blot, Immunohistochemistry, Staining, Control

Journal: Frontiers in Immunology

Article Title: MicroRNA-382 Promotes M2-Like Macrophage via the SIRP-α/STAT3 Signaling Pathway in Aristolochic Acid-Induced Renal Fibrosis

doi: 10.3389/fimmu.2022.864984

Figure Lengend Snippet: STAT3 regulates activation of macrophage in vitro , which is partly mediated by miR-382. (A) Images of immunofluorescence for p-STAT3 in DMSO+AA and STA-21+AA groups in Raw264.7 cells. Scale bars, 100 μm. (B) MFI of CD206+ macrophages in BMDMs from control, AA, DMSO+AA, and STA-21+AA groups. (C) Relative of mRNA expression of Fizz1 in BMDMs from control, AA, DMSO+AA, and STA-21+AA groups. (D–F) Relative of mRNA expression of stat3, Fizz1, and MR in Raw264.7 cells from control, AA, NC+AA, and STAT3 siRNA+ AA groups. (G) Representative immunoblot for p-STAT3 S727, p-STAT3 Y705, and STAT3 in Raw264.7 after overexpression of STAT3. GAPDH served as standard. (H) Relative of transcripts for stat3, miR-382, SIRP-α, Arg-1, Fizz1, and Ym-1 in Raw264.7 cells after STAT3 OE plasmid transfection, NC as control. (I) Images of Western blot for p-STAT3 and STAT3 in Raw264.7 cells from ctrl, AA, anti-scramble+ AA, anti-miR-382+ AA, and anti-miR-382+ STAT3 OE+ AA groups. GAPDH served as standard. (J) Abundance of miR-382 in ctrl, AA, anti-scramble+ AA, anti-miR-382+ AA, and anti-miR-382+ STAT3 OE+ AA groups. U6 served as standard. (K) MFI of CD206+ macrophages in ctrl, AA, anti-scramble+ AA, anti-miR-382+ AA, and anti-miR-382+ STAT3 OE+ AA groups. (L, M) Relative mRNA of Fizz1 and Arg-1 in ctrl, AA, anti-scramble+ AA, anti-miR-382+ AA, and anti-miR-382+ STAT3 OE+ AA groups. 18s served as standard. *P < 0.05; **P < 0.01; ANOVA.

Article Snippet: STA-21 (HY-18061, MedChemExpress, Princeton, NJ, USA) was administered intraperitoneally to animals at a dose of 0.5 mg/kg the day before AA injection (DMSO was used as control).

Techniques: Activation Assay, In Vitro, Immunofluorescence, Control, Expressing, Western Blot, Over Expression, Plasmid Preparation, Transfection