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. 2023 Feb 25;14:1078. doi: 10.1038/s41467-023-36826-0

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© The Author(s) 2023

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. 1 — A (a) Differential expression changes in lncRNA transcripts in T1-44 treated CT26 (q < 0.05) cells with respect to DMSO treatment. Transcript Per Million (TPM) expression values were normalised to the mean. Yellow, up-regulation; blue, down-regulation; ivory, minimal change. Expression data was derived from three independent experiments (each with one technical sample). (b) As above, but displaying differential expression in lncRNA transcripts in T1-44-treated colon26 tumours (p < 0.05) with respect to control. Expression data were derived from tumours collected from three mice (from one experiment). B Percentage of lncRNA genes differentially regulated at a statistically significant level in CT26 cells (left) or Colon26 tumours (right) that score as potential direct E2F1 target genes, or are associated with other potential E2F1 target genes. C RT-qPCR analysis of CT26 cells transfected with E2F1 siRNAs and treated with 1 µM T1-44 (indicated lncRNA transcripts are labelled with their ENSEMBL transcript name). An immunoblot to display input protein levels is included. SDMe was used as a marker for T1-44 activity; n = 3 independent experiments (each with three technical replicates). Results are mean values ± SD; one-way ANOVA with Tukey’s multiple comparisons test; * adjusted P value < 0.05, ** adjusted P value < 0.01, *** adjusted P value <0.001. D (a) Colon26-bearing BALB/c mice received T1-44 orally at 100 mg/kg for 19 days with respect to vehicle only control; n = 7 mice per group; experiment was performed twice; (b) Absolute tumour volume presented as a mean ± SEM, n = 7. (c) Absolute tumour volume of individual mice at day 12 (two-tailed Student’s t test; * p = 0.0143), n = 7; box and whiskers are defined as minimum, first quartile, median, third quartile, and maximum of data. (d) Relative body weight presented as a mean value ± SEM, n = 7. E Immunohistochemical staining of SDMe (a), anti-CD8 (b), anti-CD4 (c), or anti-CD163 (d) in Colon26 tumours collected at 19 days post treatment with T1-44, or at day 14 from non-treated controls. Original magnification: 20×, scale bar, 50 μm; and 63×; scale bar, 16 μm. n = 4 mice from Fig. 1D; (e) Optical density is presented as mean ± SD; two-tailed Student’s t test; n = 4 independent experiments (each performed on two separate slides); *** adjusted P value <0.001.