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. Author manuscript; available in PMC: 2016 Nov 17.
Published in final edited form as: Immunity. 2015 Nov 3;43(5):909–922. doi: 10.1016/j.immuni.2015.10.008

Figure 3. The cytosolic tail of TLR4 is not required for endocytosis.

Figure 3

(A) Schematic of three TLR4 alleles. Amino acid designations, relative to the start codon (position 1) are indicated. Ecto: extracellular domain. TM: transmembrane domain. Cyto: cytosolic tail. CD4: extracellular domain of CD4.

(B) Cells were treated with LPS or P3C for the indicated times and lysed. MyD88 was immunoprecipitated (IP) from lysates and IRAK2 and IRAK4 were detected by western blot.

(C) Indicated iBMDM lines were treated with LPS (100 ng/ml) or P3C (100 ng/ml) for 4 hours. il1b and rsad2 expression were measured by qPCR.

(D) The iBMDMs indicated were treated with LPS (1 μg/ml) for 30 min. Surface staining of TLR4 was measured by flow cytometry.

(E) The iBMDMs indicated were treated with LPS (1 μg/ml) for the times indicated. Surface staining of TLR4 was measured by flow cytometry. Line graphs represent the MFI of TLR4 surface staining at each time point, as compared to the staining pre-stimulation.

(F) The iBMDMs indicated were treated with LPS (1 μg/ml) for the times indicated. Surface staining of CD4 was measured by flow cytometry. Line graphs represent the MFI of CD4 surface staining at each time point, as compared to the staining pre-stimulation.

Error bars represent mean SEM from triplicate readings in one experiment. **,p<0.01, ns: not significant;

See also Figure S3.