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. 2022 Feb 8;11(3):589. doi: 10.3390/cells11030589

Figure 1.

Figure 1

IgE-Ag stimulation affects mitochondrial MITF Serine 73 phosphorylation. (A), RBL cells were sensitized with 100 ng/mL IgE for 2 h followed by a 0–60 min DNP challenge. Non-sensitized cells are presented as Control (Q: Quiescent). The protein levels of MITF in the whole cell lysates were determined by Western blot analysis using anti-pS73-MITF, anti-MITF and anti-GAPDH antibodies. (B), Densitometry of MITF-pS73/MITF levels in RBL cells that were activated for 0–60 min. (C), RBL and BMMCs were treated with 100 ng/mL IgE for 2 h followed by a 15 min DNP challenge and were fractionated into cytosol and mitochondria. The protein levels of MITF in both fractions were determined by Western blot analysis using anti-pS73-MITF, anti-MITF, anti-GAPDH (cytosolic marker) and anti-VDAC (mitochondrial marker) antibodies. (D), Densitometry of MITF-pS73/GAPDH and MITF-pS73/VDAC levels in RBL cells that were activated for 15 min followed by cytosolic and mitochondrial fractionation. A two-tailed Student t-test was performed for all the western blots. Results represent means ± SEMs (n = 4, **** p ≤ 0.0001; *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05; ns p > 0.05).