Fig 2: Developmental changes in the expression patterns of AQP4, FGFR3, and CD44. (A) At E96, AQP4 was expressed at LER and GER but not in hair cells and substantial sensory epithelium between inner hair cells and outer hair cells. (B) At E101, AQP4 expression was detected in some supporting cells. (C) At E115, AQP4 expression was detected in immature outer pillar cells (arrow), Hensen's cells, some GER cells, and outer sulcus cells. (D) At P0, AQP4 was expressed in inner sulcus cells, Hensen's cells, Claudius cells, and outer sulcus cells. (E) At E96, FGFR3 expression was detected in hair cells and some SOX2‐positive cells. (F, G) At E101 and E115, FGFR3 expression was detected in outer hair cells and some SOX2‐positive supporting cells. No expression was detected in inner hair cells. (H) At P0, FGFR3 expression was not detected. (I, J) No CD44 expression was detected at E96 and E101. (K) At E115, CD44 was expressed in both inner and outer pillar cells, and some Claudius cells and Hensen's cell. (L) At P0, CD44 expression was only detected in inner sulcus cells and SOX2‐negative Hensen's cells. The nuclei were counterstained with Hoechst (blue). Scale bar: 50 µm. A, B, D, E, G, H, and I: basal turn; C, F, J, K, and L: middle turn. IHC: inner hair cells, OHC, outer hair cells; IPC, inner pillar cells; OPC, outer pillar cells; HC, Hensen's cells. E96 (n = 5), E101 (n = 4), E115 (n = 4), and P0 (n = 3).

Fig 3: Disruption of the PTX3 and CD44 interaction attenuates fibrosis signalling. (A) The activity of AKT1, JNK, c‐Jun and NF‐κB (p65) in response to PTX3 treatment over the indicated time courses. Lysates from experimental cells were harvested for Western blot analysis. (B) HFL1 cells were pre‐treated with or without wortmannin (50 nM), BAY 11–7085 (1 μM) or JNK inhibitor II (7.5 μM) for 10 min and treated with or without PTX3 for 15 min. Cell lysates were immunoblotted for pAKT1, pJNK, pc‐Jun and pp65 and normalised to the respective protein levels. α‐Tubulin served as the internal control. (C) HFL1 cells were infected with shVoid or shCD44 lentiviruses and treated with PTX3 for 15 min. The activity of AKT1, JNK, c‐Jun and p65 was examined using Western blotting and normalised to the respective protein levels. α‐Tubulin was used as the internal control. (D) The indicated signalling inhibitors were pre‐treated before treatment with or without PTX3 for 24 h. Lysates from experimental cells were harvested for Western blot analysis using specific antibodies as indicated. Relative protein expression was normalised to α‐tubulin. Immunoblotting was replicated independently at least three times per experiment. (E) The migratory capacity of fibroblasts was assessed by determining the number of HFL1 cells following pre‐treatment with the indicated signalling inhibitors and then treatment with or without PTX3 for 18 h. (F) The nodule collagen formation in fibroblasts was determined by counting the nodule number in HFL1 cells following pre‐treatment with the indicated signalling inhibitors and then treatment with or without PTX3 for 24 h. Scale bars are 100 μm. All data are shown as the means ± SEM. Differences between groups were analysed using one‐way ANOVA followed by Tukey's multiple comparison test. **p < .01, ***p < .001

Fig 4: NF-κB antagonist JSH-23 abolishes the promotion of PLOD1 on malignant behavior in GBM.a, b As measured by MTS assays, JSH-23 treatment will eliminate the promotion effect of PLOD1 overexpression in PN03-GSC and PN04-GSC. c, l JSH-23 treatment regulated the protein expression of YKL40, CD44, and NF-κB downstream genes in PLOD1-overexpressed PN03-GSC and PN04-GSC as measured by western blotting. d, e Transwell assays found that the invasiveness was decreased in PLOD1 overexpression PN03-GSC and PN04-GSC after JSH-23 treatment. f, g TUNEL assays confirmed that the decreased apoptosis due to PLOD1 overexpression in PN03-GSC and PN04-GSC was significantly increased after t JSH-23 treatment. h–k JSH-23 treatment reduced the self-renewal capacity of PLOD1 overexpression PN03-GSC and PN04-GSC as measured by neurospheres formation assays and extreme limit dilution assays. Scale bar = 20 μm. All data are shown as the mean ± SD (three independent experiments). *P < 0.05; **P < 0.01; ***P < 0.001.

Fig 5: Transcriptome analyses of NSF-HA-treated IMR90 cells.a Genes differentially expressed in IMR90 cells upon treatment with cNSF-HA or fNSF-HA (HA polymer length-dependent genes). Cells were pre-incubated for 6 h with 20 μg/ml cNSF-HA, fNSF-HA, or PBS and then HA was removed and cells were exposed to 200 μM tBHP for 1 h. Cells were collected 6 h after starting the tBHP-treatment. b Enrichment of GO Term, Reactome pathway, and transcription factor protein-protein interactions (PPIs) in the HA polymer length-dependent genes. c, d Enrichment of transcription factor binding sites (TFBS) in the HA polymer length-dependent genes. White bars represents the enrichment of TFBS in all HA polymer length-dependent genes. Gray bars represents the enrichment of TFBS in HA polymer length-dependent genes that are involved in the transcriptional regulation by p53 or encoding p53-interacting proteins. Black bars represents the enrichment of TFBS in HA polymer length-dependent p53 target genes. e Differential expression levels of the HA polymer length-dependent genes (defined in the presence of tBHP) in IMR90 cells incubated for 6 h with 20 μg/ml cNSF-HA, fNSF-HA, or PBS. f The violin plot represents the distribution of p-values of a two-tailed t-test comparing the gene expression levels of the HA polymer length-dependent genes and HA-polymer length-independent genes (defined in the presence of tBHP) between cNSF-HA- and fNSF-HA-incubated IMR90 cells. Cells were incubated for 6 h with 20 μg/ml cNSF-HA or fNSF-HA without further tBHP-treatment. g Differential expression levels of the HA polymer length-dependent genes (defined in the presence of tBHP) in control and CD44-overexpressing IMR90 cells incubated for 6 h with 20 μg/ml cNSF-HA or PBS.