Extended Data Fig. 4: Systemic analysis of the impact of MDK on DCs.
(a) Mdk expression in the isogenic melanoma cell lines used in vivo to assess depletion or overexpression of this gene in Fig. 3 (n = 3 independent depletions). Data are presented as mean values +/- SEM. Statistical significances were determined by an unpaired two-tailed t-test. (b) Draining lymph nodes from B16.R2L-shCtrl and -shMdk stained with CD11c (green), CD103 (red), CD8 (purple) and DAPI (blue), and merged. Scale bar = 200 μm. Representative images of lymph nodes from three independent mice per each group. Insets highlighted in the merged are shown in Fig. 3f. (c) Total number of resident and migratory cDC2s in draining lymph nodes of B16.R2L-shCtrl implanted mice (n = 4 per condition). Statistical significances were determined by an unpaired two-tailed t-test. (d) Total number of resident and migratory cDC1s in draining lymph nodes of B16.R2-shCtrl (n = 7 mice) and -shMdk (n = 6 mice) for loss of function studies (p-value: **** <0.0001, * = 0.0336, black asterisks represent significance between resident cDC1s, and grey asterisks represent difference between migratory cDC1s). Also shown are data for B16.F1-Neg (n = 6 mice) and -Mdk (n = 6 mice) for gain of function analyses (p-value: * = 0.0163, black asterisks represent significance between resident cDC1s). Data are represented as mean values +/- SEM. Statistical significance was determined by two-way ANOVA corrected for multiple comparisons with Sidak’s multiple comparison test. (e) Percentage of circulating cDC2s in blood of mice bearing implants of B16.R2L-shCtrl (n = 16) vs shMdk (n = 19). Unpaired two-tailed t-test. (f) Percentage of circulating cDC1s, cDC2s, and pre-DCs in blood in mice bearing B16.R2-shCtrl (n = 7) or shMdk (n = 7). Statistical significance was determined by unpaired two-tailed t-test. Plots correspond to a representative experiment from two repetitions. (g) Number of cDC2s in bone marrow (femur and tibia) from mice bearing B16.R2L transduced with shCtrl (n = 5) or shMdk (n = 5). Statistical significances were determined by an unpaired two-tailed t-test. All violin plots show median values (center dash-line), quartiles (dotted-lines) and minimum and maximum values (edges of the violin), with individual values plotted.
