A. B.

Naive LLC NTC LLC sh21 Naive LLC NTC LLC sh21
Colored by phenotype:
20 CD11b+ CD11c+ CD64+ Siglec-F-
CD11b+ CD11c- CD64+ Siglec-F-
CD11b- CD11c+ CD64+ Siglec-F+
CD3+ CD4+ MHCII-
0 CD3+ CD8+ MHCII-
CD11b+ Gr-1+
CD11b+ Gr-1-
-20
CD45- MHCII+
tSNE2

CD45- MHCII-
CD19+ MHCII+
-20 0 20 -20 0 20 -20 0 20
Tumor
Unidentified
tSNE1

C. Enriched in LLC NTC D. Enriched in LLC sh21

15
*** ****
E.
* **** ** **** Distribution of Tumor Clusters
**** ***
Naive 20 Naive LLC NTC

% of events
% of events

10 LLC NTC LLC NTC

LLC sh21 LLC sh21
LLC sh21
*
Colored by phenotype:
CD45- Ki67+, #5
5 10
CD45- Ki67-, #35

11.07%
17.54%
0 0
Cluster #2 Cluster #5 Cluster #3
CD11b+ CD11c- Tumor CD11b- CD11c+
CD64+ Siglec-F- CD64+ Siglec-F+

F. Phenotypes of tumor clusters in LLC NTC Phenotypes of tumor clusters in LLC sh21
Arginase-1 IkBa Ki67 Ly6A/E MHCI MHCII PD-L1 Arginase-1 IkBa Ki67 Ly6A/E MHCI MHCII PD-L1

0 3* 0 3 0 6* 0 5 0 5 0 8 0 6 0 2.5* 0 3 0 4* 0 5 0 5 0 8 0 6

G.
Distribution of CD64+ Clusters in LLC NTC Distribution of CD64+ Clusters in LLC sh21

PD-L1 PD-L1
PD-L1 PD-L1

Colored by phenotype:
0 6
0 6 0 6 CD11b+ CD11c- CD64+ Siglec-F-, #2 0 6
CD11b- CD11c+ CD64+ Siglec-F+, #3
PD-L1 lo CD11b- CD11c+ CD64+ Siglec-F+, #4
PD-L1
CD11b+ CD11c+ CD64+ Siglec-F-, #7
PD-L1 mid
PD-L1 PD-L1 mid
PD-L1 hi PD-L1 hi
PD-L1 PD-L1
0 6

LLC NTC 0 6
LLC sh21
0 6 0 6
35.92% 37.87%