phospho ser1248 plcγ1  (Cell Signaling Technology Inc)

Journal: Microbiology Spectrum

Article Title: D-glucose uptake inhibits bovine alphaherpesvirus 1 post-binding cell process entry via inhibition of PLC-γ1 signaling in a glucose transporter 1-independent manner

doi: 10.1128/spectrum.02456-25

Figure Lengend Snippet: Glucose affects phosphorylation of PLC-γ1 at Ser1248. ( A ) MDBK cells were subjected to glucose depletion using D-glucose-free DMEM or mock depletion using high glucose DME for 12 h. The cells were then treated with D-glucose at the indicated concentrations in glucose-free DMEM for 0.5 h, after which cell lysates were prepared. The protein levels of p-PLC-γ1(Ser1248) and PLC-γ1 were detected by Western blotting. ( B ) MDBK cells were subjected to glucose depletion or mock depletion as described above. The cells were then infected with BoAHV-1 (MOI =1) for 0.5 h in the presence of D-glucose at the indicated concentrations in glucose-free DMEM. After the infection, the lysates were prepared. The protein levels of p-PLC-γ1(Ser1248) and PLC-γ1 were detected by Western blotting. β-Actin was used as a loading control and subsequent quantitative analysis. ( C ) A549 cells in 60 mm dishes were subjected to glucose depletion via culturing cells using glucose-free DMEM. Then, the cells were infected with BoAHV-1(MOI =1) for 0.5 h in the presence of D-glucose at a concentration of 25 mM in glucose-free DMEM. After the infection, the cell lysates were prepared. The protein levels of p-PLC-γ1(Ser1248) and PLC-γ1 were detected by Western blotting. The band intensity was analyzed with free software ImageJ. The results shown are the average of three independent experiments, with error bars indicating SDs. Significance was assessed with Student’s t -test (*, P < 0.05; **, P < 0.01; ns, not significant).

Article Snippet: Phospho-PLCγ1 (Ser1248) rabbit mAb (cat# 8713S), HRP-conjugated goat anti-mouse IgG (cat# 7076), and HRP-labeled goat anti-rabbit IgG (cat# 7074) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Phospho-proteomics, Western Blot, Infection, Control, Concentration Assay, Software

Journal: Microbiology Spectrum

Article Title: D-glucose uptake inhibits bovine alphaherpesvirus 1 post-binding cell process entry via inhibition of PLC-γ1 signaling in a glucose transporter 1-independent manner

doi: 10.1128/spectrum.02456-25

Figure Lengend Snippet: Schematic of putative model on the functional effects of GLUT1 and D-glucose on BoAHV-1 productive infection. GLUT1 plays an important role in the stimulation of β-catenin-dependent transcriptional effects, a potential mechanism to affect viral productive infection. D-glucose at higher concentration is capable of inhibiting virus entry partially via blocking PLC-γ1 signaling as demonstrated by reduced phosphorylation of p-PLC-γ1 at Ser1248. The graphic was created by the online program of BioGDP.com .

Article Snippet: Phospho-PLCγ1 (Ser1248) rabbit mAb (cat# 8713S), HRP-conjugated goat anti-mouse IgG (cat# 7076), and HRP-labeled goat anti-rabbit IgG (cat# 7074) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Functional Assay, Infection, Concentration Assay, Virus, Blocking Assay, Phospho-proteomics

Journal: Mediators of Inflammation

Article Title: Caffeine Acts as an Agonist of Siglec-6, Inhibits MRGPRX2-Triggered Mast Cell Degranulation and Anaphylactoid Reactions

doi: 10.1155/mi/9580121

Figure Lengend Snippet: Caffeine upregulation Siglec-6/SHP-1 signaling pathway and downregulation MRGPRX2 signaling pathway. (A) The levels of Siglec-6, p-SHP-1, SHP-1, and MRGPRX2 protein. (B–D) Quantification of Siglec-6, p-SHP-1, and MRGPRX2 protein expression. (E) The levels of PLCγ1, p-PLCγ1, IP3R, p-IP3R, ERK1/2, and p-ERK1/2 by western blot. (F–H) Quantification of p-PLC, p-IP3R, and p-ERK1/2 expression, n = 3. Significance levels were denoted as ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001.

Article Snippet: The primary antibodies used were as follows: anti-GAPDH (1:5000, 52,902) from Signalway Antibody; anti-MRGPRX2 (1:500) from lab-made; anti-SHP-1 (1:1000, # 108192-T40) from Sino-Biological; anti-phosphorylated-SHP-1 (p-SHP-1; 1:1000, #ab41436) from Abcam; anti-Siglec-6 (1:1000, ab317307) from Abcam; anti-PLCγ (1:1000, #5690) from CST; anti-phosphorylated PLCγ1 (p-PLCγ1; Ser1248) (1:1000, #8713) from CST; anti-IP3R (1:1000, #3763) from CST; anti-phosphorylated-IP3R (p-IP3R; Ser1756) (1:1000, #8548) from CST; anti-ERK1/2 (1:1000, 4695) from CST; and anti-phosphorylated ERK1/2 (p-ERK1/2; Thr202/Tyr204) (1:1000, 4370) from CST; Peroxidase AffiniPure Goat Anti-Rabbit lgG (H + L) (DY60202, 1:5000) from Deeyeebio; Peroxidase AffiniPure Goat Anti-Mouse lgG (H + L) (DY60203, 1:5000) from Deeyeebio; HRP-conjungated AffiniPure Goat Anti-Rabbit IgE (H + L) (sa00001−2, 1:10,000) from proteintech.

Techniques: Expressing, Western Blot

Journal: Mediators of Inflammation

Article Title: Caffeine Acts as an Agonist of Siglec-6, Inhibits MRGPRX2-Triggered Mast Cell Degranulation and Anaphylactoid Reactions

doi: 10.1155/mi/9580121

Figure Lengend Snippet: Caffeine targeted Siglec-6 downregulation MRGPRX2 signaling pathway. (A) The levels of p-SHP-1, SHP-1, MRGPRX2, PLCγ1, p-PLCγ1, ERK1/2, and p-ERK1/2 protein in Siglec-6 siRNA-LAD2 cells. (B) Quantification of p-SHP-1 protein expression. (C) Quantification of MRGPRX2 protein expression. (D) Quantification of p-PLCγ1 expression. (E) Quantification of p-ERK1/2 expression, n = 3. Significance levels were denoted as ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001.

Article Snippet: The primary antibodies used were as follows: anti-GAPDH (1:5000, 52,902) from Signalway Antibody; anti-MRGPRX2 (1:500) from lab-made; anti-SHP-1 (1:1000, # 108192-T40) from Sino-Biological; anti-phosphorylated-SHP-1 (p-SHP-1; 1:1000, #ab41436) from Abcam; anti-Siglec-6 (1:1000, ab317307) from Abcam; anti-PLCγ (1:1000, #5690) from CST; anti-phosphorylated PLCγ1 (p-PLCγ1; Ser1248) (1:1000, #8713) from CST; anti-IP3R (1:1000, #3763) from CST; anti-phosphorylated-IP3R (p-IP3R; Ser1756) (1:1000, #8548) from CST; anti-ERK1/2 (1:1000, 4695) from CST; and anti-phosphorylated ERK1/2 (p-ERK1/2; Thr202/Tyr204) (1:1000, 4370) from CST; Peroxidase AffiniPure Goat Anti-Rabbit lgG (H + L) (DY60202, 1:5000) from Deeyeebio; Peroxidase AffiniPure Goat Anti-Mouse lgG (H + L) (DY60203, 1:5000) from Deeyeebio; HRP-conjungated AffiniPure Goat Anti-Rabbit IgE (H + L) (sa00001−2, 1:10,000) from proteintech.

Techniques: Expressing

Journal: Mediators of Inflammation

Article Title: Caffeine Acts as an Agonist of Siglec-6, Inhibits MRGPRX2-Triggered Mast Cell Degranulation and Anaphylactoid Reactions

doi: 10.1155/mi/9580121

Figure Lengend Snippet: Caffeine inhibits PLC downstream signaling pathways. (A) The levels of PLCγ1, p-PLCγ1, ERK1/2, and p-ERK1/2 in m-3M3FBS-activated LAD2 cells. (B) Quantification of p-PLCγ1 expression. (C) Quantification of p-ERK1/2 expression, n = 3. Significance levels were denoted as ⁣ ∗ p < 0.05, ⁣ ∗∗ p < 0.01, and ⁣ ∗∗∗ p < 0.001.

Article Snippet: The primary antibodies used were as follows: anti-GAPDH (1:5000, 52,902) from Signalway Antibody; anti-MRGPRX2 (1:500) from lab-made; anti-SHP-1 (1:1000, # 108192-T40) from Sino-Biological; anti-phosphorylated-SHP-1 (p-SHP-1; 1:1000, #ab41436) from Abcam; anti-Siglec-6 (1:1000, ab317307) from Abcam; anti-PLCγ (1:1000, #5690) from CST; anti-phosphorylated PLCγ1 (p-PLCγ1; Ser1248) (1:1000, #8713) from CST; anti-IP3R (1:1000, #3763) from CST; anti-phosphorylated-IP3R (p-IP3R; Ser1756) (1:1000, #8548) from CST; anti-ERK1/2 (1:1000, 4695) from CST; and anti-phosphorylated ERK1/2 (p-ERK1/2; Thr202/Tyr204) (1:1000, 4370) from CST; Peroxidase AffiniPure Goat Anti-Rabbit lgG (H + L) (DY60202, 1:5000) from Deeyeebio; Peroxidase AffiniPure Goat Anti-Mouse lgG (H + L) (DY60203, 1:5000) from Deeyeebio; HRP-conjungated AffiniPure Goat Anti-Rabbit IgE (H + L) (sa00001−2, 1:10,000) from proteintech.

Techniques: Protein-Protein interactions, Expressing

Journal: The EMBO Journal

Article Title: The TrkC-PTPσ complex governs synapse maturation and anxiogenic avoidance via synaptic protein phosphorylation

doi: 10.1038/s44318-024-00252-9

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Rabbit anti-phospho-PLCγ1 (Ser1248) , Cell Signaling Technology , 8713S.

Techniques: Knock-In, Recombinant, Plasmid Preparation, Clone Assay, Staining, Sequencing, DC Protein Assay, Membrane, Western Blot, Electron Microscopy, Mass Spectrometry, Phospho-proteomics, Software, Fluorescence, Microscopy, Incubation