phosphorylated src src  (Cell Signaling Technology Inc)

Journal: International Journal of Surgery (London, England)

Article Title: CD36 serves as a signaling receptor and free fatty acid (FFA) transporter in dyslipidemia: the role of dapagliflozin and alirocumab in downregulating atherogenic dyslipidemia via FFA uptake inhibition—experimental study

doi: 10.1097/JS9.0000000000003451

Figure Lengend Snippet: DAPA and alirocumab inhibited lipid droplet formation and cell stress signaling depended on suppressing the expression of receptor CD36. (A) Protein expression of Fyn, * vs. other groups with different symbols (†, ‡), P < 0.0001. (B) Protein expression of Lyn, * vs. other groups with different symbols (†, ‡), P < 0.0001. (C) Protein expression of phosphorylated (p)-Src, * vs. other groups with different symbols (†, ‡), P < 0.0001. (D) Protein expression of p-MKKK, * vs. other groups with different symbols (†, ‡), P < 0.0001. (E) Protein expression of p-JNK, * vs. other groups with different symbols (†, ‡), P < 0.0001. (F) Protein expression of p-c-JUN, * vs. other groups with different symbols (†, ‡), P < 0.0001. (G) Protein expression of p-p38, * vs. other groups with different symbols (†, ‡), P < 0.0001. (H) Protein expression of p-ERK1/2, * vs. other groups with different symbols (†, ‡), P < 0.0001. (I) Protein expression of NF-κB, * vs. other groups with different symbols (†, ‡), P < 0.0001. (J) Protein expression of PPAR-γ, * vs. other groups with different symbols (†, ‡), P < 0.0001. (K) Protein expression of NOX2, * vs. other groups with different symbols (†, ‡), P < 0.0001. (L) Analytical result of ANGPTL4 activity by ELISA, * vs. other groups with different symbols (†, ‡, §, ¶, α, γ), P < 0.001. (M) Analytical result of lipoprotein lipase activity (LPLa) by ELISA, * vs. other groups with different symbols (†, ‡, §, ¶, α, γ), P < 0.0001. All statistical analyses were performed by one-way ANOVA, followed by Bonferroni multiple comparison post hoc test ( n =3 for WB in each group). Symbols (*, †, ‡, §, ¶, α, γ) indicate significance (at 0.05 level). (N–T) Illustrating the lipid droplet formation in a macrophage (gray color) in different conditions. (U) Analytical result of lipid droplet formation * vs. other groups with different symbols (†, ‡), P < 0.0001 ( n = 5). Cell grouping : F1 = RAW 264.7 cells, F2 = RAW 264.7 cells + FFA (TG), F3 = RAW 264.7 cells + FFA (TG) + DAPA (50 μM), F4 = RAW 264.7 cells + FFA (TG) + Alirocumab (200 nM), F5 = siRNA CD36 in RAW 264.7 cells + FFA, F6 = siRNA CD36 in RAW 264.7 cells + FFA + DAPA (50 μM), F7 = siRNACD36 in RAW 264.7 cells + FFA + Alirocumab (200 nM). FFA, free fatty acid.

Article Snippet: The membranes were incubated with the indicated primary antibodies [CD36 (1:4000, ABCAM), PPARr (1:1000, Cell signaling), Fyn (1:1000, Cell signaling), Lyn (1:1000, Cell signaling), phosphorylated (p)-Src (1:1000, Cell signaling), p-ERK1/2 (1:3000, Merck), NOX2 (1:4000, Sigma), p-JNK (1:3000, Abcam), p-c-Jun (1:3000, Abcam), IL-1β (1:1000, Cell signaling), p-nuclear factor keppa B (p-NF-κB) (1:1000, Cell signaling), p-MMK4 (1:4000, Abcam), p-p38 (1:4000, Sigma), ANGPTL4 (1:4000, Abcam), NOD-like receptor protein 3 (NLRP3) (1:4000, Abcam), cleaved caspase1 (1:4000, novusbio), interleukin (IL)-18 (1:3000, Abcam), and Actin (1:12 000, Merck)] for 1 hour at room temperature.

Techniques: Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: British journal of pharmacology

Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

doi: 10.1111/bph.70112

Figure Lengend Snippet: FIGURE 2 Dose-dependent effects of chronic administration of clozapine on expression of pannexin1 and phosphorylated Src in the mPFC. Rats were chronically administered with clozapine (0, 5 and 10 mg kg1 day1 for 28 days). In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH (a) and phosphorylated-Src (pSrc) per Src (b). *P < 0.05: relative to control (clozapine free) by one-way ANOVA with Scheffe's post hoc test. F-values of the chronic clozapine administration on the expression of pannexin1 and pSrc in the rat frontal cortex using one-way ANOVA were [F(3,15) = 17.3 (P < 0.01)] and [F(3,15) = 18.0 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.

Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

Techniques: Expressing, Control, Western Blot

Journal: British journal of pharmacology

Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

doi: 10.1111/bph.70112

Figure Lengend Snippet: FIGURE 6 Concentration-dependent effects of chronic administration of clozapine on expression of pannexin1 and phosphorylated Src in cultured astrocytes. (a and b) Concentration-dependent effects of chronic exposure to clozapine (3 and 10 μM for 28 days) on expression of pannexin1 and phosphorylated Src in the astroglial plasma membrane fraction, respectively. In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH or phosphorylated Src (pSrc) per Src. *P < 0.05: relative to control (clozapine free) by one- way ANOVA with Scheffe's post hoc test. F-values of the concentration-dependent effects of chronic clozapine administration on the expression of pannexin1 and pSrc using one-way ANOVA were [F(2,15) = 21.3 (P < 0.01)] and [F(2,15) = 14.9 (P < 0.01)], respectively. The lower panels indicate pseudo-gel images of capillary immunoblotting.

Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

Techniques: Concentration Assay, Expressing, Cell Culture, Clinical Proteomics, Membrane, Control, Western Blot

Journal: British journal of pharmacology

Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

doi: 10.1111/bph.70112

Figure Lengend Snippet: FIGURE 7 Interaction among clozapine and inhibitors of Src and III-mGluR on the expression of pannexin1 and phosphorylated Src in the cultured astrocytes. (a) Interaction between 10 μM PP2 (Src inhibitor) and 10 μM clozapine on pannexin1 expression in the astroglial plasma membrane. (b) Interaction between 100 μM CPPG (III-mGluR inhibitor) and 10 μM clozapine on phosphorylated-Src expression in the astroglial plasma membrane. In the upper histograms, ordinate: mean ± SD (n = 6) of the relative levels of pannexin1 (PANX) per GAPDH or phosphorylated Src (pSrc) per Src. *P < 0.05: relative to control (clozapine free) by two-way ANOVA with Scheffe's post hoc test. @: P < 0.05: relative to 10 μM clozapine exposure by two-way ANOVA with Scheffe's post hoc test. F-values of the interaction between 10 μM PP2 and chronic exposure to 10 μM clozapine on the expression of pannexin1 using two-way ANOVA were [Fclozapine(1,20) = 55.5 (P < 0.01), FPP2(1,20) = 0.2 (P > 0.05), Fclozapine*PP2(1,20) = 1.1 (P > 0.05)]. F- values of the interaction between 100 μM CPPG and chronic exposure to 10 μM clozapine on the expression of pSrc using two-way ANOVA were [Fclozapine(1,20) = 56.0 (P < 0.01), FCPPG(1,20) = 27.7 (P < 0.01), Fclozapine*CPPG(1,20) = 0.4 (P > 0.05)]. The lower panels indicate pseudo-gel images of capillary immunoblotting.

Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

Techniques: Expressing, Cell Culture, Clinical Proteomics, Membrane, Control, Western Blot

Journal: British journal of pharmacology

Article Title: Exploring the pathophysiology underlying clozapine-induced enhancement of glutamatergic transmission through L-glutamate and D-serine release associated with pannexin1 hemichannels.

doi: 10.1111/bph.70112

Figure Lengend Snippet: FIGURE 9 Effects of chronic administration of L-BAIBA on Src signalling in the cultured astrocytes. (a and b) Interaction among chronic exposure to 30 μM L-BAIBA for 28 days and inhibitors of III-mGluR (100 μM CPPG) and NMDAR (1 μM MK801), respectively. In the upper side histogram, ordinate: mean ± SD (n = 6) of the relative levels of phosphorylated-Src (pSrc) per Src. *P < 0.05: relative to control (L-BAIBA free) by two-way ANOVA with Scheffe's post hoc test. @: P < 0.05: relative to 30 μM L-BAIBA exposure alone. F-values of the interaction between 100 μM CPPG and chronic exposure to 30 μM L-BAIBA on the expression of pSrc using two-way ANOVA were [FL-BAIBA(1,20) = 27.6 (P < 0.01), FCPPG(1,20) = 59.3 (P < 0.01), FL-BAIBA*CPPG(1,20) = 2.7 (P > 0.05)]. F-values of the interaction between 1 μM MK801 and chronic exposure to 30 μM L-BAIBA on the expression of pSrc using two-way ANOVA were [FL-BAIBA(1,20) = 46.3 (P < 0.01), FMK801(1,20) = 1.0 (P > 0.05), FL-

Article Snippet: Primary antibodies against GAPDH (NB300-327, RRID:AB_10001915, 1:300; Novus Biologicals, Littleton, CO, USA), pannexin1 (12595-1-AP, RRID:AB_ 2159186, 1:100; Proteintech, Rosemont, IL, USA), Src (2109, RRID: AB_2106059,1:50; Cell Signalling Technology, Danvers, MA, USA) and phosphorylated Src (pSrc; MAB2685, RRID:AB_3657975, 1 μg ml 1, R&D Systems, Minneapolis, MN, USA) were diluted in an antibody diluent (Immuno Shot Platinum; CosmoBio, Tokyo, Japan).

Techniques: Cell Culture, Control, Expressing