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. Author manuscript; available in PMC: 2020 Jul 21.
Published in final edited form as: Cell Rep. 2020 Jun 16;31(11):107771. doi: 10.1016/j.celrep.2020.107771

Figure 2. STING N153S Reduces Numbers of LTi Cells in Fetal Tissues as Well as Their Accumulation in Developing LN Anlagen.

Figure 2.

In (A) and (B), leukocytes were harvested from the fetal gut on E16.5–E18.5 and analyzed by flow cytometry. In (C)–(F), frozen sections of cervical LN anlagen of E18.5 fetuses were analyzed by widefield fluorescence microscopy.

(A) Representative FACS dot plots of Lin−CD45+CD127+cKIT+ cells isolated from the fetal gut. Numbers indicate the percent of events in each gate.

(B) Total number α4β7+RORγT+ LTi cells in the fetal gut on E16.5–E18.5. Data represent the mean of n = 20–23 fetuses per genotype pooled from three independent experiments.

(C) Cervical LN anlagen sections were stained with DAPI and with antibodies against VCAM-1 (left panel) and CD4 (middle panel). Scale bar: 40 μm.

(D) Quantitation of CD4 staining intensity relative to the size of LN anlagen, defined as the total area of VCAM-1 and CD4 staining.

(E) Quantitation of VCAM-1 staining intensity relative to the size of LN anlagen.

(F) Quantitation of the size of LN anlagen, based on merged VCAM-1 and CD4 staining of WT and STING N153S cervical LN anlagen on E18.5.

Quantitation is from n = 7–9 cervical LN anlagen per genotype from two independent experiments. FACS data were analyzed by unpaired t test, and immunofluorescence data were analyzed by Mann-Whitney U test. *p < 0.05; ****p < 0.0001.