ZFIN Image: Finotto et al., 2023, Figure EV3

Image

Figure Caption

Figure EV3 GSCC‐specific morphometrics and dynamics of the tumor and its microenvironment in 3D over time

A

Number of zebrafish embryos used for the time‐lapse movies that were generated at 1 and 5 dpi.

B

Mean tumor volume over time, during 1 dpi movies (left) and 5 dpi movies (right) (n = 10 (BT333), 12 and 11 (BT569; 1 and 5 dpi), 10 and 9 (CME037; 1 and 5 dpi), 6 (CME038), 8 and 4 (LBT001; 1 and 5 dpi), 22 and 18 (LBT003; 1 and 5 dpi), 12 and 9 (LBT070; 1 and 5 dpi), 15 and 11 (LBT123; 1 and 5 dpi) zebrafish embryos, see also panel A).

C

Tumor volume at the start of 1 dpi time‐lapse movies (n = 10 (BT333), 12 (BT569), 10 (CME037), 6 (CME038), 8 (LBT001), 22 (LBT003), 12 (LBT070), 15 (LBT123); P = < 1.0e‐15 (BT333), 0.0001 (BT569), 0.0017 (CME037), 3.1e‐5 (CME038), < 1.0e‐15 (LBT001), 0.0002 (LBT070), 0.1542 (LBT123)).

D

Tumor volume at the start of 5 dpi time‐lapse movies (n = 10 (BT333), 11 (BT569), 9 (CME037), 6 (CME038), 4 (LBT001), 18 (LBT003), 9 (LBT070), 11 (LBT123); P = 0.0009 (BT333), 0.0179 (BT569), 0.0237 (CME037), 0.0095 (CME038), 0.0538 (LBT001), 4.7e‐5 (LBT070), 0.0003 (LBT123)).

E

Representative maximum intensity projections of a z stack of the head region of a Tg(mpeg1:mCherryF)^ump2; Tg(kdrl:lynEYFP)^md77 zebrafish embryo with a GFP‐labeled LBT070 tumor, at 37.5, 38, 38.5 and 39 hpi to illustrate phagocytosis of a GBM tumor cell by a round GAM (indicated by arrows). GBM tumor cells are shown in green, GAMs in red, and blood vessels in blue. Scale bars: 50 μm.

F, G

Trend line of median GAM distance to the tumor over time in 1 dpi (F) and 5 dpi (G) movies for all GSCCs (n = 43,857 (F) and 16,639 (G) GAMs; P = 6.0e‐7 (F) and 0.7071 (G)).

H

Boxplot of distance of round and ramified GAMs to the tumor for all GSCCs, at the start of 1 and 5 dpi time‐lapse movies (n = 1,821 and 1,429 (round; 1 and 5 dpi), 420 and 1,008 (ramified; 1 and 5 dpi) GAMs; P < 6.4e‐7 (1 dpi) and < 2.4e‐5 (5 dpi); boxes stand for 50% of the data and minima/maxima are indicated by the line ends).

I

Boxplot of GAM distance to the tumor of round GAMs within 30 μm of the tumor, at the start of 1 and 5 dpi time‐lapse movies, ranked by increasing median distance at 1 dpi (n = 325 and 146 (LBT003; 1 and 5 dpi), 273 and 44 (LBT123; 1 and 5 dpi), 134 and 105 (BT333; 1 and 5 dpi), 141 and 49 (BT569; 1 and 5 dpi); 137 and 48 (LBT070; 1 and 5 dpi), 74 and 56 (CME037; 1 and 5 dpi), 26 and 26 (CME038; 1 and 5 dpi), 61 and 11 (LBT001; 1 and 5 dpi); boxes stand for 50% of the data and minima/maxima are indicated by the line ends).

J

Boxplot of GAM distance to the tumor of ramified GAMs within 30 μm of the tumor, at the start of 1 and 5 dpi time‐lapse movies, ranked by increasing median distance at 1 dpi (n = 63 and 51 (LBT003; 1 and 5 dpi), 26 and 26 (LBT123; 1 and 5 dpi), 38 and 21 (BT569; 1 and 5 dpi), 13 and 12 (CME038; 1 and 5 dpi); 29 and 13 (LBT070; 1 and 5 dpi), 25 and 86 (BT333; 1 and 5 dpi), 18 and 30 (CME037; 1 and 5 dpi), 15 and 3 (LBT001; 1 and 5 dpi); boxes stand for 50% of the data and minima/maxima are indicated by the line ends).

Data information: Data describe biological replicates, i.e. individual zebrafish embryos. The xenograft experiment was replicated once for CME038, twice for BT333, BT569, CME037, LBT001, three times for LBT070 and LBT123, and four times for LBT003. In (C and D), data are presented as mean ± SD. In (C), the P‐values were calculated by Welch's ANOVA test, followed by Dunnett's T3 multiple comparisons correction. In (D), the P‐values were calculated by Kruskal–Wallis test, followed by Dunn's multiple comparisons correction. In (F and G), the P‐values were calculated by the Mann–Kendall trend test. In (H), the P‐values were calculated by Welch's ANOVA test, followed by Games‐Howell's multiple comparisons correction. ns ≥ 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Source data are available online for this figure.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ EMBO Mol. Med.