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. 2023 May 1;53(7):2250056. doi: 10.1002/eji.202250056

Figure 4.

Figure 4

TLR4 Y672 and Y749 are not essential for LPS‐inducible TLR4 internalization. Tlr4−/− BMM were reconstituted with WT mouse TLR4, the indicated TLR4 mutants or empty vector (EV). (A) The median fluorescence intensities of the TLR4 signal in unstimulated cells were compared between GFP–ve (untransduced) and GFP+ve (transduced) populations. (B) Cells were treated for the indicated time with 10 ng/mL LPS and assessed for cell‐surface TLR4 via flow cytometry. Within the GFP+ve population, the median fluorescence intensities of each TLR4‐expressing population are presented following the subtraction of the median fluorescence intensity of EV (TLR4−/− ) samples. (A & B) Data (mean ± SEM, n = 3–4) are combined from three to four independent experiments. Statistical analyses were performed using a repeated measures two‐way ANOVA (A) or a mixed‐effect analysis (B), followed by Bonferroni's multiple comparisons test (NS: nonsignificant, *p < 0.05, ***p < 0.001, ****p < 0.0001).