Journal: Signal Transduction and Targeted Therapy

Article Title: Prostaglandin E 2 -driven dedifferentiation of Schwann cells leads to perineural invasion in pancreatic ductal adenocarcinoma

doi: 10.1038/s41392-026-02648-x

Figure Lengend Snippet: SCs are enriched in the PNI-positive region. a Schematic diagram of the workflow; some of the graphical elements were created wtith BioGDP. b Division of perineural invasion regions (PNI-region: PNI, PNI-near, PNI-mid, PNI-away) and nonperineural invasion regions (NPNI: NPNI, NPNI-near, NPNI-mid, NPNI-away) in representative spatial transcriptomic images of PDAC PNI samples, with corresponding representative HE-stained images of spot points (right) (n = 4). N: Nerve, T: Tumor. Scale bar: 500 μm. c GSVA of the PNI region and the NPNI region. d Correlations between different cell types and the expression levels of marker genes. e Cell type distribution scatter plot based on the UMAP dimensionality reduction algorithm; f UMAP plot of SC subsets in pancreatic cancer tissues. g Feature score of nomyelinating SCs at the single-cell level in each of the two SC subsets ( p = 0.025). h Representative images of RCTD single-cell mapping results based on PNI regions and NPNI regions (n = 4); i Stacked bar charts of different PNI regions and NPNI regions, showing the proportions of various cell types in different samples. j , k Representative mIHC images of human normal pancreatic tissues, PDAC-PNI- tissues, and PDAC-PNI+ tissues stained with PGP9.5, SOX2, c-Jun, and MUC1 antibodies. Scale bar: 100 μm (left), 50 μm (right). The ratios of SOX2+ and c-Jun+ positive cells in normal pancreatic tissues, PDAC-PNI- tissues, and PDAC-PNI+ tissues ( n = 5). All the results are presented as the means ± SDs. Each data point in the bar graphs represents an individual sample. Statistical significance was determined by a two-tailed unpaired t test (Fig. 1g) and one-way analysis of variance followed by Tukey’s HSD post hoc test (Fig. 1j). **** p < 0.0001

Article Snippet: For mIHC staining, formalin-fixed sections were stained with antibodies against PGP9.5 (1:200, Abways, CY6722), MUC1 (1:200, BOSTER, BM4548), SOX2 (1:200, Diagbio, db16528), c-Jun (1:200, Selleck, F0168), and p75NTR (1:200, ABclonal, A11169) according to the manufacturer’s instructions for the multicolor immunofluorescence kit (Zolgene, PR-004507) and imaged with a KFbio Pathology Imaging System (KF-FL-005).

Techniques: Staining, Expressing, Marker, Single Cell, Two Tailed Test

Journal: Signal Transduction and Targeted Therapy

Article Title: Prostaglandin E 2 -driven dedifferentiation of Schwann cells leads to perineural invasion in pancreatic ductal adenocarcinoma

doi: 10.1038/s41392-026-02648-x

Figure Lengend Snippet: Pancreatic cancer cells promote SC dedifferentiation. a Schematic diagram of the coculture system of PDAC cells and SCs. b Venn diagram of upregulated pathways in RSC96 (+PANC-1) cells and RSC96 (+BxPC-3) cells compared with RSC96 cells ( n = 3). c , e Transwell assays were used to evaluate the effects of PANC-1 and BxPC-3 cells on the migration of sNF96.2 cells ( n = 6). Scale bar: 100 μm. d Microscopy images showing the morphological changes in sNF96.2 cells induced by PANC-1 and BxPC-3 cells. Scale bars: 50 μm (upper panels) and 20 μm (lower panels). f – j Western blotting was performed to detect the protein expression levels of dedifferentiation markers (p75NTR, c-Jun, SOX2, N-cadherin, and GDNF) in sNF96.2 cells after coculture with PANC-1 or BxPC-3 cells ( n = 3). k , m Transwell assays were used to assess the effects of PANC-1 and BxPC-3 cells on the migration of RSC96 cells ( n = 6). Scale bar: 100 μm. l Microscopy images showing the morphological changes in RSC96 cells induced by PANC-1 and BxPC-3 cells. Scale bars: 50 μm (upper panels) and 20 μm (lower panels). n – r Western blotting was conducted to determine the protein expression levels of dedifferentiation markers (p75NTR, c-Jun, SOX2, N-cadherin, and GDNF) in RSC96 cells after coculture with PANC-1 or BxPC-3 cells (n = 3). Relative expression levels were quantified as p75NTR/GAPDH, c-Jun/GAPDH and GDNF/GAPDH; since the molecular weight of SOX2 (~35 kDa) is close to that of GAPDH (~36 kDa), SOX2/β-actin was used for relative expression quantification. All the results are presented as the means ± SDs. Each data point in the bar graphs represents an individual sample. Statistical significance was determined by one-way analysis of variance followed by Tukey’s HSD post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: For mIHC staining, formalin-fixed sections were stained with antibodies against PGP9.5 (1:200, Abways, CY6722), MUC1 (1:200, BOSTER, BM4548), SOX2 (1:200, Diagbio, db16528), c-Jun (1:200, Selleck, F0168), and p75NTR (1:200, ABclonal, A11169) according to the manufacturer’s instructions for the multicolor immunofluorescence kit (Zolgene, PR-004507) and imaged with a KFbio Pathology Imaging System (KF-FL-005).

Techniques: Migration, Microscopy, Western Blot, Expressing, Molecular Weight

Journal: Signal Transduction and Targeted Therapy

Article Title: Prostaglandin E 2 -driven dedifferentiation of Schwann cells leads to perineural invasion in pancreatic ductal adenocarcinoma

doi: 10.1038/s41392-026-02648-x

Figure Lengend Snippet: Pancreatic cancer-derived PGE 2 induces SC activation. a Representative IF images of DiI and PTGER1/2/4 in RSC96 cells. Scale bars: 20 μm (left), 5 μm (right). b Representative F-actin IF images of RSC96 and sNF96.2 cells after treatment with different doses of PGE 2 (0 nM, 0.1 nM, 1 nM, or 10 nM). Scale bar: 50 μm. c , e – g Protein expression levels of p75NTR, c-Jun, and SOX2 in RSC96 cells treated with different doses of PGE 2 (0 nM, 0.1 nM, 1 nM, or 10 nM). Relative expression levels were quantified as p75NTR/GAPDH and c-Jun/GAPDH; since the molecular weight of SOX2 (~35 kDa) is close to that of GAPDH (~36 kDa), the relative expression of SOX2/β-actin was quantified ( n = 3). d , h – j Representative IF images of p75NTR, c-Jun, and SOX2 in RSC96 cells after PGE 2 intervention. Scale bar: 100 μm. Statistics of the relative fluorescence intensity between the control and treatment groups are shown ( n = 3). k , o - q Protein expression levels of p75NTR, c-Jun, and SOX2 in RSC96 cells (RSC96-PANC-1 coculture system) following CAY10526 treatment and PGE 2 supplementation ( n = 3). l , r–t Protein expression levels of p75NTR, c-Jun, and SOX2 in RSC96 cells (RSC96-BxPC-3 coculture system) following CAY10526 treatment and PGE 2 supplementation ( n = 3). m Effects of CAY10526 treatment and PGE 2 supplementation on 3D cocultures of RSC96 cells with HPNE, PANC-1 or BxPC-3 cells ( n = 3).Scale bar: 200 μm. +B/P/H: +BxPC-3/PANC-1/HPNE, + DMSO:+BxPC-3/PANC-1/HPNE + DMSO, + CAY:+BxPC-3/PANC-1/HPNE + DMSO + CAY10526, + CAY + PGE 2 :+BxPC-3/PANC-1/HPNE + DMSO + CAY10526 + PGE 2 . n , u , v effects of CAY10526 treatment and PGE 2 supplementation on PANC-1/BxPC-3-DRG cocultures, along with quantification of the DRG growth index ( n = 3). Scale bar: 500 μm. +B/P: +BxPC-3/PANC-1, +DMSO: +BxPC-3/PANC-1 + DMSO, +CAY: +BxPC-3/PANC-1 + DMSO + CAY10526, +CAY+. PGE 2 : +BxPC-3/PANC-1 + DMSO + CAY10526 + PGE 2. All the results are presented as the means ± SDs. Each data point in the bar graphs represents an individual sample. Statistical significance was determined by one-way analysis of variance followed by Tukey’s HSD post hoc test (for e – g , o – t , and u , v ) and a two-tailed unpaired t test (for Fig. 5h-j). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: For mIHC staining, formalin-fixed sections were stained with antibodies against PGP9.5 (1:200, Abways, CY6722), MUC1 (1:200, BOSTER, BM4548), SOX2 (1:200, Diagbio, db16528), c-Jun (1:200, Selleck, F0168), and p75NTR (1:200, ABclonal, A11169) according to the manufacturer’s instructions for the multicolor immunofluorescence kit (Zolgene, PR-004507) and imaged with a KFbio Pathology Imaging System (KF-FL-005).

Techniques: Derivative Assay, Activation Assay, Expressing, Molecular Weight, Fluorescence, Control, Two Tailed Test

Journal: PloS one

Article Title: A rare myelin protein zero (MPZ) variant alters enhancer activity in vitro and in vivo.

doi: 10.1371/journal.pone.0014346

Figure Lengend Snippet: Figure 1. Analysis of MCSs in the genomic region encompassing MPZ. (A) Representation of the MPZ gene and its flanking regions from the UCSC Human Genome Browser. From top to bottom, tracks are shown displaying information about MPZ-MCSs, MPZ exons, overall multi-species sequence conservation, and pair-wise sequence conservation between human and five other vertebrate species. MPZ-MCS3 within MPZ intron 1 is indicated by an arrow. (B) Sequence chromatograms for a region within MPZ-MCS3 are shown for an unaffected individual (control) and patient BAB1733. Two monomeric consensus SOX10-binding sequences in a head-to-head configuration are indicated by red highlights and horizontal arrows. The position of the c.126-1086T.A variant (heterozygous in the patient) is indicated with a vertical arrow. (C, D) Multi-sequence alignments of the consensus SOX10-binding sequences within MPZ-MCS3 (highlighted in red) are shown containing sequences from the indicated species, as generated by MultiPipMaker (C) or available on the UCSC Human Genome Browser (D). The position of c.126-1086T in each alignment is indicated by a vertical arrow; dots indicate bases that are identical to the human reference sequence and dashes represent gaps within the alignments. doi:10.1371/journal.pone.0014346.g001

Article Snippet: The SOX10 expression construct was obtained from Origene (Rockville, MD).

Techniques: Sequencing, Control, Binding Assay, Variant Assay, Generated

Journal: PloS one

Article Title: A rare myelin protein zero (MPZ) variant alters enhancer activity in vitro and in vivo.

doi: 10.1371/journal.pone.0014346

Figure Lengend Snippet: Figure 2. Functional characterization of the c.126-1086T.A variant in MPZ-MCS3. (A) Genomic segments spanning each MPZ MCS (MPZ- MCS1, MPZ-MCS2, MPZ-MCS3, and MPZ-MCS4) were tested for their ability to direct luciferase reporter-gene expression in cultured Schwann (S16) cells compared to a control vector (pE1b). Error bars indicate standard deviation. (B) Wild-type (Wt) and c.126-1086T.A variant (Var) forms of MPZ-MCS3 were tested for their ability to direct luciferase reporter-gene expression in the forward (blue) and reverse (red) orientations in cultured Schwann (S16) cells compared to a control vector (pE1b). Error bars indicate standard deviation. (C) Similar experiments as described in B were performed with cultured oligodendrocytes (OliNeu) cells. (D) Wild-type (Wt) and c.126-1086T.A (Var) forms of MPZ-MCS3 were tested for their responsiveness to SOX10 via luciferase reporter-gene expression in SOX10-negative MN1 cells, in the forward orientation compared to a control vector (pE1b). The amount of SOX10 expression vector co-transfected is shown. Error bars indicate standard deviation and p-values were calculated using the student’s t-test. (E) Electrophoretic mobility shift analysis (EMSA) of wild-type and c.126-1086T.A alleles. ‘Probes’ (radiolabeled oligonucleotides) and ‘competitors’ (unlabeled oligonucleotides) corresponding to a 25-bp fragment including the wild-type (Wt) and c.126-1086T.A variant-containing (Var) consensus SOX10-binding sequence in MPZ-MCS3 (see Fig. 1B) were incubated with S16 nuclear extracts and subjected to electrophoresis (see Materials and Methods). Each assay used the same amount of probe coupled with varying amounts of competitor, as shown (‘none’ indicates no competitor; * and ** indicate a probe to competitor ratio of 1:10 and 1:100, respectively). Arrows denote specific nuclear proteins bound to the probe. doi:10.1371/journal.pone.0014346.g002

Article Snippet: The SOX10 expression construct was obtained from Origene (Rockville, MD).

Techniques: Functional Assay, Variant Assay, Luciferase, Gene Expression, Cell Culture, Control, Plasmid Preparation, Standard Deviation, Expressing, Transfection, Electrophoretic Mobility Shift Assay, Binding Assay, Sequencing, Incubation, Electrophoresis