Journal: Journal of Pharmacopuncture

Article Title: Suppression of Macrophage Migration and M2 Polarization by an Ethanolic Extract of Scutellaria baicalensis Root

doi: 10.3831/KPI.2025.28.2.144

Figure Lengend Snippet: Effects of ESB on STAT3 phosphorylation in RAW 264.7 cells. RAW 264.7 mouse macrophages were stimulated with IL-6 (100 ng/mL, A, B) or IL-4 (100 ng/mL, C, D) in the presence or absence of ESB (50 and 100 μg/mL). After 24 h, the levels of phosphorylated and total STAT3 and STAT6 were analyzed using Western blot analysis. Actin served as a loading control. Representative blot images from three independent experiments are shown (A, C). The ratio of phosphorylated to total protein was quantified by measuring band intensity (B, D). Data are presented as the mean ± SD of three independent experiments. Statistical significance was evaluated using one-way ANOVA with Tukey’s post hoc test ( ## p < 0.01, ### p < 0.001 versus untreated control; ns, not significant; *p < 0.05, ***p < 0.01, ***p < 0.001 versus cells treated with IL-6 or IL-4). ESB, ethanolic extract of Scutellaria baicalensis root; IL, interleukin; STAT3, signal transducer and activator of transcription 3.

Article Snippet: Total STAT3, total STAT6, and actin antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Molecular immunology

Article Title: Decreasing effects of protein kinase inhibitors on the expression of NOS2 and inflammatory cytokines and on phagocytosis in rat peritoneal macrophages is partly related to repolarization.

doi: 10.1016/j.molimm.2022.11.002

Figure Lengend Snippet: Fig. 6. Phosphorylation of STAT3 and STAT6 proteins in rat macrophages. pSTAT3/STAT3 ratio (a) and protein bands (b., pSTAT3, STAT3 and GAPDH) are shown. JAK/ STAT kinase inhibitors and IL-4 were added to macro phages from casein-treated rats during 24 h culture. IL-4 treatment caused significant (*p < 0.05), while AT and CYT caused strongly significant (***p < 0.001) decrease in phosphorylation of STAT3. Polyclonal anti-pSTAT3 anti body (rabbit, R & D Systems, 1:1000), monoclonal anti- STAT3 antibody (mouse, R & D Systems, 1:2000) and anti-GAPDH (R& D Systems, 1:3000) were used as primary, while HRPO-conjugated goat anti-rabbit (R & D Systems, 1:2000) and goat anti-mouse (R & D Systems, 1:3000) were used as secondary antibodies. c.) Results of cell-based ELISA experiments for pSTAT6 and STAT6 are shown. * *p < 0.01 between res 0′ and cas 0′; §p < 0.05 between casein 0′ and 24 h; ###p < 0.001 for IL-4, AT and CYT treatments compared to casein 24 h as control. Polyclonal anti-pSTAT6 antibody (rabbit, R & D Systems, 1:200), monoclonal anti-STAT6 antibody (mouse, R & D Systems, 1:200) and monoclonal anti-GAPDH (mouse, R & D Sys tems, 1:300) were used as primary, while HRPO- conjugated goat anti-rabbit (R & D Systems, 1:800) and goat anti-mouse (R & D Systems, 1:800) were used as secondary antibodies.

Article Snippet: Fixed cells were then treated with 50 μl rabbit polyclonal anti-phospho-STAT6 antibody (1:200, R & D Systems, MN USA), 50 μl mouse monoclonal anti-STAT6 antibody (1:200, R & D Systems) and 50 μl mouse monoclonal anti-GAPDH antibody (1:300, R & D Systems) for overnight at 4 ◦C.

Techniques: Phospho-proteomics, In-Cell ELISA, Control