Phospho-Src (Tyr419) Antibody - #AF3162

Fig. 4 Galectin-3 inhibits TREM2/DAP12 receptor complex to suppress Src/Syk signaling pathway and altered macrophage to an M2-like phenotype. (A, B) 293T cells were transfected with plasmids as shown, and anti-HA (B) or anti-Flag (B) antibodies were employed for exogenous CO-IP experiments. (C-E) 293T cells were transfected with plasmids as shown and anti-HA (C), anti-Flag (D), or anti-Myc (E) antibodies were employed for exogenous CO-IP experiments, respectively. (F, G) After 24 h of LLC CM treatment with the addition of GB1107 (5 µM), and 2 h of stimulation with LPS (1 ng/mL), the phosphorylation levels of Src and Syk in indicated time point were analyzed by western blot (F). The gray values of p-Syk and p-Src protein bands were analyzed by Image J software, and the relative gray values were standardized to the gray values of Syk and Src (G). (H, I) After 24 h of LLC CM treatment with the addition of GB1107 (5 µM), and 2 h of stimulation with LPS (1 ng/mL), the phosphorylation levels of Src and Syk in WT or TREM2 KO BMDMs were analyzed by western blot (H). The gray values of p-Syk and p-Src protein bands were analyzed by Image J image analyses software, and the relative gray values were standardized to the gray values of Syk and Src (I). (J, K) After 24 h of LLC CM treatment with the addition of rm galectin-3 (200 ng/ml), and 2 h of stimulation with LPS (1 ng/mL), the phosphorylation levels of Src and Syk in WT or TREM2 KO BMDMs were analyzed by western blot (J). The gray values of p-Syk and p-Src protein bands were analyzed by Image J image analyses software, and the relative gray values were standardized to the gray values of Syk and Src (K). (L) Phagocytosis assay between WT or TREM2 KO BMDMs which were pretreated with LLC CM supplemented with GB1107 (5 µM) for 24 h and LLC were detected by flow cytometry. (M) Following a 24 h-treatment with LLC CM supplemented with GB1107 (5 µM), the F-actin polarization of RAW264.7 cells blocked with anti-TREM2 antibody was observed by confocal microscopy. Scale bars, 5 μm. Quantitative statistics of F-actin polarization were analyzed by Image J image analyses software. (N-P) After 24 h of LLC CM which was supplemented with GB1107 (5 µM) treatment, the transcription levels of Ccr2(N), M2-like macrophage markers (CD206, Arg1) (O) and M1-like macrophage markers (Nos2, TNFα) (P) in WT or TREM2 KO BMDMs were detected by RT-qPCR assay. Data represent mean ± SD from three experiments. *, P 

Fig. 4 Matrine was a non-ATP-competitive inhibitor of Src kinase. A The effect of matrine on Src kinase activity in cancer cells, including HT-29, MCF7, A549, BxPC-3, SKOV3, and HeLa, were determined according to the instructions of Src kinase activity detection kit. Results are representative of three experiments (*P 

Fig. 4 MTSS1 inactivated Src and activated Wnt/β-catenin signaling in stromal progenitor cells. ST2 cells were transfected with MTSS1 expression construct (A, D) or MTSS1 siRNA (B, E) or Src expression construct (F) and their control. The protein levels of MTSS1, phosphorylated, and total Src were detected in the cells 3 days after transfection using Western blotting (A, B). Co-IP was performed and endogenous MTSS1 and LRP6 were detected with Western blotting (C). The protein levels of the major components of Wnt/β-catenin signaling were detected in the cells 3 days after transfection using Western blotting (D–F). Values represent mean ± SD, n = 3. Student’s t test was conducted in (A, D, F). One-way ANOVA was conducted in (B, E) followed by Tukey test for post hoc comparisons. *Significant vs. vector or control siRNA, p 

Figure 8 FOR increases AhR expression and inhibits the phosphorylation level of Src. The present results are displayed as the mean ± SEM of triple parallel measurements. #(P < 0.01) is significantly different from control group; **(P < 0.01) is significantly different from LPS group.

Fig. 5. |Jinfukang inhibited the cell migration via Integrin/Src axis in lung cancer cells. Cells were incubated with Jinfukang (100 μg/mL), ATN-161 (10 μmol/L) or a combination of both for 24 h. A Western blot assay was used to measure the protein expression levels of Integrin β1, Src, and p-Src in CTC-TJH-01 (A) and H1975 (B)cells. GAPDH was used as an internal standard.

FIGURE 5 | Cyclin G2 inhibits the FAK-SRC-STAT pathway in vitro and in vivo. (A) The protein expressions along with quantification of FAK, P-FAK, SRC, P-SRC, STAT3, P-STAT3, Bcl-2, c-Myc, and MMP9 in the FAK-SRC-STAT pathway. (B–D) The relative mRNA expressions of Bcl-2, c-Myc and MMP9. (E) Analysis of MMP9 secretion and its quantification in SCC-9 cells. (F, G) Immunohistochemical staining and analysis of the expression of cyclin G2, p-FAK, p-SRC and p-STAT3. Scale bar = 100 µm. *p < 0.05, **p < 0.01, ***p < 0.001 vs. vector.

FIGURE 3 Molecular profiling for iAs-induced changes in Caco-2 cells. (A) Protein levels of claudin 3 and cofilin were determined by Western blot analysis. (B) Transcript levels of IL-6, TNF-α, and MMP13 were determined by RT-qPCR, with GAPDH as an internal control. (C) The levels of EGF in the culture supernatant were examined by ELISA. (D) A marked increase in the phosphorylated levels of EGFR, Src, AKT, and p38 was shown by Western blot analysis. RT-qPCR (E) and Western blot analysis (F) results showed the down-regulation of HTRA1 expression caused by chronic iAs exposure. Data were representatives of at least three independent experiments, shown as mean ± SD. The significance threshold for Student's t-test: