Journal: Nature Communications

Article Title: Quantification of ligand and mutation-induced bias in EGFR phosphorylation in direct response to ligand binding

doi: 10.1038/s41467-023-42926-8

Figure Lengend Snippet: A Raw dose response curves for Y1068 and Y1173 phosphorylation per EGFR molecule, for the ligands EGF, TGFα, and epiregulin. Each point represents the ratio of either anti-pY1068 or anti-pY1173 fluorescence and EGFR-mTurq fluorescence for one individual vesicle. Each curve contains ~1000 to ~3000 data points (single vesicles). B Bias plots. Shown are means (symbols) and standard errors (often smaller than symbols). The epiregulin points diverge from the EGF and TGFα points. In total, data are from 11,570 single vesicles over 25 independent experiments. C Bias coefficients and standard errors. Epiregulin is biased toward Y1173 phosphorylation as compared to EGF and TGFα. Ordinary one-way ANOVA, followed by Tukey’s test, was used to determine statistical significance. The p values are adjusted for multiple comparisons.

Article Snippet: Y1173 phosphorylation was detected using 50 nM AlexaF488-labeled anti-pY1173 EGFR antibody (NBP1-44893AF488, Novus Biologicals) .

Techniques: Phospho-proteomics, Fluorescence

Journal: Nature Communications

Article Title: Quantification of ligand and mutation-induced bias in EGFR phosphorylation in direct response to ligand binding

doi: 10.1038/s41467-023-42926-8

Figure Lengend Snippet: A Single vesicle dose response curves for Y1068 and Y1173 phosphorylation per EGFR molecule, for EGF, TGFα, and epiregulin. Each point represents the ratio of either anti-pY1068 or anti-pY1173 fluorescence and EGFR-mTurq fluorescence for one individual vesicle. Each curve contains ~1000 to ~1800 data points. B Bias plots. Shown are means (symbols) and standard errors (often smaller than symbols). In total, data are from 8009 vesicles in 23 independent experiments. C Bias coefficients and their standard errors. Ordinary one-way ANOVA, followed by Tukey’s test, was used to determine statistical significance. The p values are adjusted for multiple comparisons.

Article Snippet: Y1173 phosphorylation was detected using 50 nM AlexaF488-labeled anti-pY1173 EGFR antibody (NBP1-44893AF488, Novus Biologicals) .

Techniques: Phospho-proteomics, Fluorescence

Journal: Drug Design, Development and Therapy

Article Title: Blockade of the P2Y2 Receptor Attenuates Alcoholic Liver Inflammation by Targeting the EGFR-ERK1/2 Signaling Pathway

doi: 10.2147/DDDT.S346376

Figure Lengend Snippet: Hepatoprotective effect of suramin was associated with inhibiting EGFR-ERK1/2 signaling pathway. ( A – D ) The protein expression of P2Y2R, TNF-α and IL-1β in the liver. ( E and F ) The protein expression levels of EGFR, p-EGFR, ERK1/2 and p-ERK1/2 in the liver. *** P < 0.001 compared to control group. ## P < 0.01, ### P < 0.001 compared to Alcohol group.

Article Snippet: Antibodies against total ERK, phospho-ERK1/2 (p-ERK1/2), total EGFR and phospho-EGFR (p-EGFR) were obtained from Elabscience (Wuhan, China).

Techniques: Expressing, Control

Journal: Drug Design, Development and Therapy

Article Title: Blockade of the P2Y2 Receptor Attenuates Alcoholic Liver Inflammation by Targeting the EGFR-ERK1/2 Signaling Pathway

doi: 10.2147/DDDT.S346376

Figure Lengend Snippet: ERK1/2 inhibition mitigated alcohol-induced AML-12 cell inflammation. ( A and B ) The effect of UTP (1.89 μM) and suramin (200 μM) on alcohol-induced phosphorylation of ERK1/2 in AML-12 cell. ( E and F ) The effect of the EGFR antagonist AG1478 (3 nmol/L) on alcohol-induced phosphorylation of ERK1/2 in AML-12 cell. ( G and H ) The effect of the ERK1/2 antagonist U0126 (65 nmol/L) on alcohol-induced phosphorylation of ERK1/2 in AML-12 cell. ( I – K ) The effect of the ERK1/2 antagonist U0126 (65 nmol/L) on alcohol-induced protein and mRNA expression of TNF-α and IL-1β in AML-12 cell. ** P < 0.01, *** P < 0.001 compared to the control group. # P < 0.05, ## P < 0.01 compared to the alcohol group, && P < 0.01 compared to the alcohol plus UTP group. ( C and D ) The effect of P2Y2R silencing and UTP on alcohol-induced phosphorylation of ERK1/2 in AML-12 cell. ** P < 0.01 compared to the control group. ## P < 0.01 compared to the negative control group.

Article Snippet: Antibodies against total ERK, phospho-ERK1/2 (p-ERK1/2), total EGFR and phospho-EGFR (p-EGFR) were obtained from Elabscience (Wuhan, China).

Techniques: Inhibition, Phospho-proteomics, Expressing, Control, Negative Control

Journal: Drug Design, Development and Therapy

Article Title: Blockade of the P2Y2 Receptor Attenuates Alcoholic Liver Inflammation by Targeting the EGFR-ERK1/2 Signaling Pathway

doi: 10.2147/DDDT.S346376

Figure Lengend Snippet: Blockade of the P2Y2 receptor attenuates alcoholic liver inflammation by targeting the EGFR-ERK1/2 signaling pathway.

Article Snippet: Antibodies against total ERK, phospho-ERK1/2 (p-ERK1/2), total EGFR and phospho-EGFR (p-EGFR) were obtained from Elabscience (Wuhan, China).

Techniques: