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. 2022 Feb 17;13:952. doi: 10.1038/s41467-022-28638-5

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — A Examples of R variant M. abscessus-tdTomato lesions in DAPI-stained cryosections from a 14 dpi TgBAC(tnfa:GFP)^pd1028 adult zebrafish. i. Cellular lesion, ii. Necrotic granuloma. Arrow indicates necrotic granuloma, M. abscessus-tdTomato is coloured magenta, tnfa promoter induction is marked in green. Scale bars indicate 100 μm. Images are representative of two biological replicates of 3 animals per replicate. B Examples of S variant M. abscessus-tdTomato lesions in DAPI-stained cryosections from a 28 dpi TgBAC(tnfa:GFP)^pd1028 adult zebrafish. Arrows indicate necrotic granulomas, M. abscessus-tdTomato is coloured magenta, tnfa promoter induction is marked in green. Scale bar indicates 100 μm. Images are representative of two biological replicates of 3 animals per replicate. C i. Examples of tnfa promoter activity in 14 dpi TgBAC(tnfa:GFP)^pd1028 adult zebrafish infected with M. abscessus-tdTomato and treated with dexamethasone as indicated. Arrows indicate necrotic granulomas, M. abscessus-tdTomato is coloured magenta, tnfa promoter induction is marked in green. Scale bars indicate 100 μm. Control images are representative of 3 animals per M. abscessus variant, dexamethasone images are representative of 2 animals per M. abscessus variant. ii. Quantification of GFP pixels as a function of M. abscessus-tdTomato fluorescence in TgBAC(tnfa:GFP)^pd1028 adult zebrafish. Data are a single experimental replicate, each point represents a single granuloma, total animals per group: Control R = 3, Dexamethasone R = 2, Control S = 3, Dexamethasone S = 3. Statistical testing by two-sided ANOVA. D Enumeration of CFUs from adult zebrafish infected with either the R or the S variant of M. abscessus and treated with dexamethasone. Each point represents a single animal and data are representative of 2 biological replicates. Total number of animals: Control R = 4, Dexamethasone R = 3, Control S = 5, Dexamethasone S = 6. Statistical testing by two-sided Student’s t test within each of the independent R and S experiments. Data are presented as mean values ± SD. Source data are provided as a Source Data file.