Fig. 2: Modelling HPV16 E6E7 integration and C. trachomatis coinfection in ectocervical organoids.

a, b Time-lapse live imaging of hCEcto (a) and hCEcto E6E7 (b) organoids infected with GFP-expressing C. trachomatis (GFP-Ctr). Representative time-lapse microscopic images at indicated time points after Ctr infection. Arrows highlight the progression of Ctr infection from basal cells to daughter cells in the basal and differentiated layers. c, d Representative images of hCEcto (c) and hCEcto E6E7 (d) organoids at 1 and 5dpi with GFP-Ctr of three biological replicates. e, f Infectivity of lysates from Ctr infected hCEcto (e) and hCEcto E6E7 (f) organoids at 1 and 5dpi. Shown is the mean ± SD of two replicates representative of three biological replicates. g, h Primary infection of Ctr infected hCEcto and hCEcto E6E7 cells (g) and inclusions size (h). Whiskers are min–max, box is 25–75th percentile, lines are median values. n = biological replicates. i Infectivity of Ctr infected hCEcto and hCEcto E6E7 2D stem cells lysates at 48 hpi. Shown is the mean ± SD of three technical replicates. j, k Transmission electron micrographs of Ctr infected hCEcto (j) and hCEcto E6E7 (k) 2D stem cells at 24hpi. Inclusion contains different development forms of Ctr (RB, reticulate bodies; EB, elementary bodies; IB, intermediate stage; AB, aberrant bodies). Shown are representative images from three biological replicates. l Percentage of total EBs, RBs, IBs, and ABs in inclusions, representative of three biological replicates. Data are presented as violin plot min–max, lines are 25 and 75th percentile, bold line is median. Confocal images of hCEcto (m–o) and hCEcto E6E7 (p–r) organoids at 5dpi immunolabeled for Ki67 (m, p); KRT5 (n, q), p63 (o, r) and Ctr-MOMP, Nuclei in blue. Representative images of three biological replicates. s, t Quantification of the number of Ki67 positive cells from confocal images of uninfected and Ctr infected hCEcto and hCEcto E6E7 2D stem cells at 48hpi (s) and organoids at 5dpi (t). Whiskers are min–max, box is 25–75th percentile, lines are median values. (g–i, l, s, t) Statistical significance was calculated using two-sided t test, P-values are indicated. Source data are provided as a Source Data file.