Fig. 7: Dynamics of MOR C-terminus phosphorylation. | Communications Biology

Fig. 7: Dynamics of MOR C-terminus phosphorylation.

From: High-throughput screening and validation of antibodies against synaptic proteins to explore opioid signaling dynamics

Fig. 7

Neuro2A and Neuro2A PKCα-KO cells were treated with PMA (a, d), morphine (b, e), or fentanyl (c, f) for different time intervals, and probed with anti-phospho S363 MOR (AVG710P-B7) (a–c) and anti-phospho S375/T376 MOR (AVG713-B5) (d–f) antibodies. Images were acquired with the InCell microscope, and the fluorescence intensity was obtained using the CellProfiler 3.1.5 software. Data are mean ± SE of two experiments n = 150–3000 cells. *p value < 0.01; ***p value < 0.001; ****p value < 0.0001; One-way ANOVA. g–i Neuro2A and Neuro2A PKCα-KO cells were treated with morphine and fentanyl for 1 h and probed using anti-phospho S375/T376 MOR (g), anti-phospho T370 MOR (AVG711P-B6) (h), and anti-phospho T379 MOR (AVG712P-G10) (i) antibodies. Bar graph, each dot represents one cell. Lines are mean ± SE of two experiments, number of counted cells are indicated. The percentage analysis used the mean of the integrated fluorescence intensity from the non-treated Neuro2A cells as 100%. NS non-significant; *p value < 0.05; ***p value < 0.001; ****p value < 0.0001; One-way ANOVA.

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