Supplementary Figure 2: Stromal expansion in TDLNs. | Nature Immunology

Supplementary Figure 2: Stromal expansion in TDLNs.

From: Tumor-induced stromal reprogramming drives lymph node transformation

Supplementary Figure 2

(a) Flow cytometry measurement of FRC number in TDLN. Red line denotes baseline FRC percentage/LN. (b) Confocal images of FRC networks in NDLNs (top) and day 11 TDLNs (bottom) LNs stained for podoplanin (red) and collagen I (blue). (c) Confocal Airyscans of conduit side and end views from NDLNs (top) and day 11 TDLNs (bottom) stained for podoplanin (green) ER-TR7 (blue) and CCL21 (red). (d) Confocal Airyscans of conduit side and end views from NDLNs (top) and day 11 TDLNs (bottom) stained for podoplanin (green) ER-TR7 (red) and collagen I (blue). (e) Quantification of network branch length. (f-h) Scatterplots showing the log2 fold expression change for all genes in the array (x axis), ranked according to increasing expression value (y axis); NDLNs vs. day 4 TDLNs (f), NDLNs vs. day 11 TDLNs (g), day 4 TDLNs vs. day 11 TDLNs (h). (i) Primary eigenvectors for Principal Component Analysis. The vast majority of change in the data (93.4%) are contained within the first 2 eigenvalues. Scale bars (b) 50 µm, (c) 0.487 µm (NDLN end), 0.291 µm (NDLN side), 0.588 µm (TDLN end), 0.873 µm (TDLN side), (d) 0.437 µm, 0.441 µm, 0.256 µm, 0.328 µm (NDLN end) and 0.454 µm (NDLN side); 0.363 µm, 0.481 µm, 0.47 µm, 0.363 µm (TDLN end) and 0.55 µm (TDLN side). Each symbol represents an individual FOV (e). Small horizontal lines indicate mean ± s.e.m. *P <0.05 (two-tailed unpaired t-test (a)). Data represent two independent experiments n=4 NDLNs, n= 6 TDLNs (mean ± s.e.m. (a)); or three independent experiments in female C57BL/6 mice, n=6 NDLNs and n=4 TDLNs.

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