Journal: Research

Article Title: Mapping Immune-Inflammatory Niches on Zirconia Bone Implants: Single-Cell Transcriptomic Profiling

doi: 10.34133/research.1162

Figure Lengend Snippet: Ti and ZrO 2 implants induce early osseointegration and immune inflammation. (A) Transcriptional levels of osteogenesis-related and immune-inflammatory genes. Osteogenic genes: Runx2 , Osterix , and Tgfb1 . Immune-inflammatory genes: Il1b , Mmp9 , and Il23a . (B) Receiver operating characteristic (ROC) analysis based on bulk RNA-seq data. AUC, area under the curve. (C) Protein expression levels of osteogenesis-related and immune-inflammatory markers. Osteogenic proteins: RUNX2, OSX, TGFB1, pSMAD2/SMAD2, and pSMAD3/SMAD3. Immune-inflammatory proteins: IL1B, MMP9, and IL23A. (D) Statistical analysis of protein expression levels. Statistical significance was determined using one-way analysis of variance. RUNX2, RUNX family transcription factor 2; OSX, osterix; TGFB1, transforming growth factor beta 1; SMAD2, SMAD family member 2; SMAD3, SMAD family member 3; IL1B, interleukin 1 beta; MMP9, matrix metallopeptidase 9; IL23A, interleukin 23 subunit alpha.

Article Snippet: The membranes were blocked with 5% nonfat milk for 1 h, followed by overnight incubation at 4 °C with the following primary antibodies: RUNX2 (Bioss, bs-1134R, 1:1,000, China), OSX (Bioss, bs-25532R, 1:1,000, China), pSMAD2 (Bioss, bs-3419R, 1:1,000, China), pSMAD3 (Bioss, bs-3425R, 1:1,000, China), IL1B (Bioss, bs-0812R, 1:1,000, China), MMP9 (Bioss, bsm-54040R, 1:3,000, China), IL23A (Bioss, bs-1193R, 1:1,000, China), NOS2 (Bioss, bs-0162R, 1:2,000, China), CD206 (Bioss, bsm-55604R, 1:1,000, China), TGFB1 (Proteintech, 81746-2-RR, 1:1,000, China), SMAD2 (Proteintech, 12570-1-AP, 1:3,000, China), and SMAD3 (Proteintech, 30130-1-AP, 1:2,000, China).

Techniques: RNA Sequencing, Expressing

Journal: bioRxiv

Article Title: Restoration of Keratinocyte Homeostasis Drives Resolution of Skin Inflammation

doi: 10.64898/2026.03.04.708224

Figure Lengend Snippet: a, Volcano plots depicting global differential gene expression across disease and restoration states. Left: KC-Tie2 versus C57BL/6 skin under control conditions, defining disease-associated transcriptional changes. Middle: KC-Tie2 skin following ω-3 PUFA exposure versus KC-Tie2 control skin, illustrating transcriptional changes accompanying disease normalization. Right: Subset of disease-associated genes that are significantly modulated in the opposite direction during ω-3 PUFA-mediated disease resolution. Genes were filtered using |log₂ fold change| ≥ 1.5 and FDR < 0.05; non-significant genes are shown in grey. Selected genes with large effect sizes are labeled. The transgene Tek and the epithelial regulators Serpinb3b , Slc25a5 , and Cacna2d1 are highlighted and examined further in subsequent figures. Summary statistics for all differential expression contrasts, including DEG counts and directionality, are provided in Supplementary Table 3. b, Expression of inflammatory mediators ( Il23a , Il17a , Tnf , Il22 , Il1f6 , and Il1f5 ) in dorsal skin from C57BL/6 and KC-Tie2 mice under control or ω-3 PUFA exposure. Data are shown as individual biological replicates with mean ± s.e.m. Statistical significance was determined by two-way ANOVA with Tukey’s post hoc testing; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant. c, KEGG pathway enrichment analysis of genes increased (upper) or decreased (lower) in KC-Tie2 skin relative to C57BL/6 controls (Disease Effect). Enrichment of cardiomyopathy and contractility-related annotations reflect overrepresentation of actin-myosin and calcium-handling genes that are shared across tissues. d, KEGG pathway enrichment analysis of genes decreased (upper) or increased (lower) during ω-3 PUFA-mediated restoration in KC-Tie2 skin (Disease Resolution), demonstrating bidirectional normalization of disease-associated pathways. Dot size represents gene count; color denotes odds ratio; the x-axis indicates −log₁₀(FDR). Full gene lists and enrichment statistics are provided in Supplementary Tables 4-7. e, Density plots of log₂ fold-change distributions for genes associated with basal, differentiated, and keratinized epidermal compartments. KC-Tie2 versus C57BL/6 skin under control conditions demonstrates disease-associated transcriptional shifts across epidermal layers (upper). ω-3 PUFA-exposed KC-Tie2 skin versus control-fed KC-Tie2 skin shows redistribution of effect sizes toward baseline (lower), indicating that both disease-associated dysregulation and its normalization are concentrated within epidermal gene programs. f, Schematic summary integrating transcriptional analyses across disease and resolution states. Chronic inflamed skin is characterized by coordinated activation of IL-17-driven inflammatory signaling, keratinocyte stress responses, disrupted lipid and fatty acid metabolism, and altered calcium and cytoskeletal programs. Disease resolution is associated with suppression of inflammatory modules alongside reactivation of epidermal differentiation, barrier formation, lipid metabolism, calcium signaling, and structural homeostasis, consistent with keratinocyte-centered, systems-level transcriptional reprogramming.

Article Snippet: Mouse primers (ThermoFisher Scientific) used in this study were: Il23a , Mm01160011_g1; Il17a , Mm00439618_m1; Il17c, Mm00521397_m1, Ccl20 , Mm00444228_m1; S100a8 , Mm00495696_g1; Tnf , Mm00443260_g1, Cacna2d1 , Mm00486607_m1; Serpinb3b , Mm03032256_uH; Slc25a5 , Mm00846873_g1; and Gapdh , Mm99999915_g1.

Techniques: Gene Expression, Control, Labeling, Quantitative Proteomics, Expressing, Activation Assay

Journal: bioRxiv

Article Title: Restoration of Keratinocyte Homeostasis Drives Resolution of Skin Inflammation

doi: 10.64898/2026.03.04.708224

Figure Lengend Snippet: a, Experimental schematic for topical siRNA-mediated gene silencing in KC-Tie2 mice. Non-targeting control siRNA or gene-specific siRNAs targeting Cacna2d1 , Serpinb3b , or Slc25a5 were applied topically to ear skin at the indicated time points, with tissue collection on day 14. b, Representative hematoxylin and eosin–stained sections of ear skin following treatment with control siRNA or gene-specific siRNAs. c, Heatmap summarizing changes in epidermal acanthosis and expression of IL-23/IL-17-axis-associated genes ( Il23a , Ccl20 , S100a8 , Il17a , Tnf , Il17c ) following gene-specific knockdown. Values are shown as log₂ fold change relative to control siRNA-treated skin. Acanthosis is coded as increased or decreased relative to control. d, Correlation heatmap relating each targeted gene to transcriptional programs identified by bulk RNA-seq pathway analyses, including fatty acid and lipid metabolism, epidermal barrier and junctional organization, IL-17-associated inflammatory signaling, epidermal differentiation and cornified envelope formation, cholesterol and sterol biosynthesis, and neutrophil activation/degranulation. Quantitative log₂ fold-change values underlying the cytokine and alarmin heatmap in c are provided in Supplementary Table 29.

Article Snippet: Mouse primers (ThermoFisher Scientific) used in this study were: Il23a , Mm01160011_g1; Il17a , Mm00439618_m1; Il17c, Mm00521397_m1, Ccl20 , Mm00444228_m1; S100a8 , Mm00495696_g1; Tnf , Mm00443260_g1, Cacna2d1 , Mm00486607_m1; Serpinb3b , Mm03032256_uH; Slc25a5 , Mm00846873_g1; and Gapdh , Mm99999915_g1.

Techniques: Control, Staining, Expressing, Knockdown, RNA Sequencing, Activation Assay

Journal: bioRxiv

Article Title: A Novel human IL-23A Overexpressing Mouse Model of Systemic Lupus Erythematosus

doi: 10.1101/2023.11.08.566163

Figure Lengend Snippet: (A) Schematic representation of the human IL23A transgene . (B) human IL23A mRNA detected by PCR and (C) human IL23A protein detected by immunohistochemistry in the skin, cervical lymph nodes, spleen, lung and kidney of TghIL23A mice. Scale bar 200μm and 100μm.

Article Snippet: Tissue paraffin sections were stained with H&E for histopathological evaluation or with biotinylated anti-mouse-IgG (Vector; BA-9200) for the detection of immune complex deposits and anti-human IL23A (OriGene; AM20386PU-N) for the detection of human IL23A protein in various tissues.

Techniques: Immunohistochemistry

Journal: bioRxiv

Article Title: A Novel human IL-23A Overexpressing Mouse Model of Systemic Lupus Erythematosus

doi: 10.1101/2023.11.08.566163

Figure Lengend Snippet: Reduced skin (upper panel) and ear (lower panel) clinical (A) and histopathological signs (B), as well as reduced LN/BW and Spleen/BW ratios (C) , urine protein levels (D) IgG circulating levels (E) and glomerular IgG deposits (F) following a 10-week-long treatment of TghIL23A mice with anti-human IL23A (Guselkumab:GUS). Scale bar 20μm. Data are presented as mean ± SEM; * P < 0.04; ** P < 0.0092; *** P = 0.0002; **** P < 0.0001.

Article Snippet: Tissue paraffin sections were stained with H&E for histopathological evaluation or with biotinylated anti-mouse-IgG (Vector; BA-9200) for the detection of immune complex deposits and anti-human IL23A (OriGene; AM20386PU-N) for the detection of human IL23A protein in various tissues.

Techniques:

Journal: Oncology letters

Article Title: microRNA-23a promotes cell growth and metastasis in gastric cancer via targeting SPRY2-mediated ERK signaling.

doi: 10.3892/ol.2018.8374

Figure Lengend Snippet: Figure 5. AGS cells were co‑transfected with miR‑23a inhibitor and SPRY2‑specific siRNA, miR‑23a in+siSPRY2, or co‑transfected with miR‑23a inhibitor and non‑specific siRNA, miR‑23a in+siNC. Following this, (A) reverse transcription‑quantitative polymerase chain reaction and (B) western blot analysis were used to examine the mRNA and protein expression of SPRY2. (C) An MTT, (D) wound healing and (E) Transwell assay were conducted to examine cell proliferation, migration and invasion, respectively. **P<0.01 vs. miR‑23a in+siNC. The magnification for wound healing assay image was x40, for Transwell assay was x200. si, small interfering; miR, microRNA; NC, negative control; SPRY, sprouty homolog 2; in, inhibitor.

Article Snippet: AGS cells were transfected with 100 nM of scrambled miR mimic (miR-NC; Guangzhou Fulengen Co., Ltd., Guangzhou, China.), miR-23a mimic (miR-23a; Guangzhou Fulengen Co., Ltd.), miR-23a inhibitor (Fulengen), negative control inhibitor (NC inhibitor; Guangzhou Fulengen Co., Ltd.), co-transfected with miR‐23a inhibitor and non‐specific siRNA (miR‐23a in+siNC; OriGene Technologies, Inc., Beijing, China.) or co-transfected with miR‐23a inhibitor and SPRY2‐specific siRNA (miR‐23a in+siSPRY2; OriGene Technologies, Inc.).

Techniques: Polymerase Chain Reaction, Western Blot, Expressing, Transwell Assay, Migration, Wound Healing Assay, Negative Control

Journal: Oncology letters

Article Title: microRNA-23a promotes cell growth and metastasis in gastric cancer via targeting SPRY2-mediated ERK signaling.

doi: 10.3892/ol.2018.8374

Figure Lengend Snippet: Figure 6. (A) Western blot analysis was used to examine the levels of the p‑ERK and t‑ERK protein levels in AGS cells transfected with miR‑23a inhibitor or negative control (NC) inhibitor. **P<0.01 vs. NC inhibitor. (B) Western blot analysis was used to examine levels of the p‑ERK protein and t‑ERK protein levels in AGS cells co‑transfected with miR‑23a inhibitor and SPRY2‑specific siRNA (miR‑23a in+siSPRY2), or co‑transfected with miR‑23a inhibitor and non‑specific siRNA (miR‑23a in+siNC). **P<0.01 vs. miR‑23a in+siNC. ERK, extracellular signal‑regulated kinases; NC, negative control; miR, microRNA; si, small interfering; SPRY2, sprouty homolog 2; p, phospho; t, total; in, inhibitor.

Article Snippet: AGS cells were transfected with 100 nM of scrambled miR mimic (miR-NC; Guangzhou Fulengen Co., Ltd., Guangzhou, China.), miR-23a mimic (miR-23a; Guangzhou Fulengen Co., Ltd.), miR-23a inhibitor (Fulengen), negative control inhibitor (NC inhibitor; Guangzhou Fulengen Co., Ltd.), co-transfected with miR‐23a inhibitor and non‐specific siRNA (miR‐23a in+siNC; OriGene Technologies, Inc., Beijing, China.) or co-transfected with miR‐23a inhibitor and SPRY2‐specific siRNA (miR‐23a in+siSPRY2; OriGene Technologies, Inc.).

Techniques: Western Blot, Transfection, Negative Control

Journal: PloS one

Article Title: Interleukin 23 promotes hepatocellular carcinoma metastasis via NF-kappa B induced matrix metalloproteinase 9 expression.

doi: 10.1371/journal.pone.0046264

Figure Lengend Snippet: Figure 3. Knocking down IL-23 expression inhibits cell motility. (A), (B) IL-23 p40 expression was efficiently silenced by IL-23p40 shRNA as determined by RT-PCR, qPCR (A) and western blot (B) in MHCC-97L cell. 18S and b-actin were used as loading control respectively. (C) and (D) Compared to vector control the migration (C) and invasion (D) ability of IL-23 silenced cells were greatly decreased in MHCC-97L cell. *, P,0.05. doi:10.1371/journal.pone.0046264.g003

Article Snippet: RNA interference (RNAi) assay MHCC-97L cells were transfected with a pGFP-RS plasmid encoding IL-23p40 shRNA (Origene, Rockville MD) or a scramble control pGFP-V-RS using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instruction.

Techniques: Expressing, shRNA, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Plasmid Preparation, Migration

Journal: PloS one

Article Title: Interleukin 23 promotes hepatocellular carcinoma metastasis via NF-kappa B induced matrix metalloproteinase 9 expression.

doi: 10.1371/journal.pone.0046264

Figure Lengend Snippet: Figure 5. IL-17A promots IL-23expression in HCC. (A) Expression of IL-23 was detected by western blot analysis in HCC cells treated with or without rhIL-17A (50 ng/mL) for 24 hours. (B) Transcriptional expressions of both subunits of IL-23 were compared by qPCR between cells treated with and without rhIL-17A (50 ng/mL) for 12 hours. *, P,0.05. (C) Transcriptional expression of IL-23p40 was detected by qPCR in PLC8024 and MHCC-97L cells with different treatment. Control: without rhIL-17A treatment; rhIL-17A: treated with rhIL-17A (50 ng/mL); Helenalin+rhIL-17A: treated with helenalin (0.l mM) and rhIL-17A (50 ng/mL). *, P,0.05. doi:10.1371/journal.pone.0046264.g005

Article Snippet: RNA interference (RNAi) assay MHCC-97L cells were transfected with a pGFP-RS plasmid encoding IL-23p40 shRNA (Origene, Rockville MD) or a scramble control pGFP-V-RS using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instruction.

Techniques: Expressing, Western Blot, Control

Journal: Oncotarget

Article Title: Investigation of IL-23 (p19, p40) and IL-23R identifies nuclear expression of IL-23 p19 as a favorable prognostic factor in colorectal cancer: a retrospective multicenter study of 675 patients

doi:

Figure Lengend Snippet: A) Kaplan-Meier survival curve of test cohort showing survival time benefit of nuclear IL-23p19 positive patients particularly at early time points and B) a multivariable analysis of this cohort at 4-years follow-up. Stratification by CD8+ T-cell counts shows C) survival time benefit in patients with nuclear IL-23p19 positive cancers in CD8+ high setting with D) no benefit in CD8+ low setting.

Article Snippet: A myc-tagged IL-23p19 (IL-23A) pCMV6 expression plasmid was purchased from Origene (Labforce, Nunningen, Switzerland) and 4 μG of this plasmid were transfected into 293T HEK cells using calcium phosphate technique.

Techniques:

Journal: Oncotarget

Article Title: Investigation of IL-23 (p19, p40) and IL-23R identifies nuclear expression of IL-23 p19 as a favorable prognostic factor in colorectal cancer: a retrospective multicenter study of 675 patients

doi:

Figure Lengend Snippet: Nuclear IL-23p19 in cohort 1 stratified by CD8+ low and high groups

Article Snippet: A myc-tagged IL-23p19 (IL-23A) pCMV6 expression plasmid was purchased from Origene (Labforce, Nunningen, Switzerland) and 4 μG of this plasmid were transfected into 293T HEK cells using calcium phosphate technique.

Techniques: Mutagenesis

Journal: Oncotarget

Article Title: Investigation of IL-23 (p19, p40) and IL-23R identifies nuclear expression of IL-23 p19 as a favorable prognostic factor in colorectal cancer: a retrospective multicenter study of 675 patients

doi:

Figure Lengend Snippet: A) Kaplan-Meier survival curve of low and high CD8+ T-cell counts in patients not receiving a preoperative therapy (n=100). B) Kaplan-Meier curve showing combined nuclear IL-23p19 and CD8+ T-cell counts and effect on overall survival. Nuclear IL-23p19 positive/CD8+ high patients have a 100% survival. Effect of IL-23p19 negative/CD8+ high or IL23-p19 positive/CD8+ low has a more unfavorable intermediate effect while IL-23p19 negative/CD8+ low patients have the most limited benefit. C) Multivariable analysis showing the independent prognostic effect of the combined IL-23p19 positive/CD8+ effects on overall survival.

Article Snippet: A myc-tagged IL-23p19 (IL-23A) pCMV6 expression plasmid was purchased from Origene (Labforce, Nunningen, Switzerland) and 4 μG of this plasmid were transfected into 293T HEK cells using calcium phosphate technique.

Techniques:

Journal: Oncotarget

Article Title: Investigation of IL-23 (p19, p40) and IL-23R identifies nuclear expression of IL-23 p19 as a favorable prognostic factor in colorectal cancer: a retrospective multicenter study of 675 patients

doi:

Figure Lengend Snippet: Immunoblot showing both nuclear and cytoplasmic fractions for IL-23p19 (IL-23A) and appropriate controls.

Article Snippet: A myc-tagged IL-23p19 (IL-23A) pCMV6 expression plasmid was purchased from Origene (Labforce, Nunningen, Switzerland) and 4 μG of this plasmid were transfected into 293T HEK cells using calcium phosphate technique.

Techniques: Western Blot

Journal: Oncotarget

Article Title: Investigation of IL-23 (p19, p40) and IL-23R identifies nuclear expression of IL-23 p19 as a favorable prognostic factor in colorectal cancer: a retrospective multicenter study of 675 patients

doi:

Figure Lengend Snippet: Representative photomicrographs of positive/high (A, C, E, G) and negative/low expression (B, D, F, H) of A-B: IL-23 poly; C-D: IL-23p40; E-F: IL-23R and G -H: IL-23p19. 100 × magnification.

Article Snippet: A myc-tagged IL-23p19 (IL-23A) pCMV6 expression plasmid was purchased from Origene (Labforce, Nunningen, Switzerland) and 4 μG of this plasmid were transfected into 293T HEK cells using calcium phosphate technique.

Techniques: Expressing

Journal: Oncotarget

Article Title: Investigation of IL-23 (p19, p40) and IL-23R identifies nuclear expression of IL-23 p19 as a favorable prognostic factor in colorectal cancer: a retrospective multicenter study of 675 patients

doi:

Figure Lengend Snippet: Association of nuclear IL-23p19 expression with clinicopathological data of Cohort 1

Article Snippet: A myc-tagged IL-23p19 (IL-23A) pCMV6 expression plasmid was purchased from Origene (Labforce, Nunningen, Switzerland) and 4 μG of this plasmid were transfected into 293T HEK cells using calcium phosphate technique.

Techniques: Expressing, Mutagenesis