Journal: JID Innovations

Article Title: CIT tumor lines: A series of immunogenic murine cutaneous squamous cell carcinoma cell lines derived from chemical carcinogenesis

doi: 10.1016/j.xjidi.2026.100477

Figure Lengend Snippet: CIT lines express identifiable neoantigens whose cognate CD8+ T cells are responsive to immunotherapy. ( a ) Selection criteria for CIT6 mutations predicted to generate neoantigens. (b) Predicted peptide log affinity ratio (affinity ratio is defined as the ratio between predicted binding affinity of mutant versus wild-type sequence) and gene expression level of mutations predicted to be top CIT6 candidate neoantigens, shown for H2-Dq and H2-Kq alleles. Affinity predictions were generated using the NetH2Pan algorithm. (c) Schematic of IFNγ ELISpot assay. For screening of CD8+ T cell and CD4+ T cell responses, CD8+ or CD4+ T cells were isolated from the spleens of tumor-bearing mice and co-cultured with total splenocytes from a healthy tumor-naïve mouse. (d) Representative IFNγ ELISpot images of the assay performed with CD8+ T cells from the spleen of CIT6-bearing mice cultured with 160ug/ml of Brinp3 peptide CPAFLPCTV (top row) and negative control (no peptide, bottom row). (e) Quantification of average IFNγ ELISpot spot counts for CD8+ T cells from the spleen of CIT6-bearing mice cultured with different concentrations of Brinp3 peptide CPAFLPCTV. Each colored line represents data points from a different mouse (n = 4 mice). (f) Representative IFNγ ELISpot images of the assay performed with CD8+ T cells from the spleen of CIT6-bearing mice cultured with 160ug/ml of Ttll4 peptide VPPSSLLPL (top row) and negative control (no peptide, bottom row). (g) Quantification of average IFNγ ELISpot spot counts for CD8+ T cells from the spleen of CIT6-bearing mice cultured with different concentrations of Ttll4 peptide VPPSSLLPL. Each colored line represents data points from a different mouse (n = 4 mice). (h) Spot counts from ELISpot assays performed with CD4+ T cells isolated from the spleens of CIT6-bearing mice and cultured with splenocytes from tumor-naïve mice loaded with increasing concentrations of Brinp3 and Ttll4 peptides. Each colored line represents data points from a different mouse (n = 4 mice). (i) Spot counts from ELISpot assays performed with total splenocytes from CIT6-bearing mice cultured with increasing concentrations of Brinp3 and Ttll4 peptides. Each colored line represents data points from a different mouse (n = 10 – 11 mice). (j) Schematic of ICI treatment regimen and ELISpot analysis. (k) Average spot counts of ICI-treated splenocytes cultured with 80 ug/ml of Ttll4 peptide vs negative control. Each line represents data points from a different mouse (n = 6 mice). Statistical analysis in panels e, g, h, i, and k was done by a paired t- test. ∗ P < .05, ∗∗ P < .01. Error bars in panels e, g, h and i represent mean and SD. CIT, carcinogen-induced tumor; ICI, Immune checkpoint inhibitor.

Article Snippet: Eight-week-old wild-type FVB/N mice were purchased from Jackson Laboratories.

Techniques: Immunopeptidomics, Selection, Binding Assay, Mutagenesis, Sequencing, Gene Expression, Generated, Enzyme-linked Immunospot, Isolation, Cell Culture, Negative Control

Journal: iScience

Article Title: MAP3K1 integrates RhoA/ROCK signaling to regulate epithelial morphogenesis

doi: 10.1016/j.isci.2026.115853

Figure Lengend Snippet: S1P activates the MAP3K1-RhoA signaling network (A) Western blot and (B) densitometric analysis of phosphoproteins in wild type and Map3k1 −/− keratinocytes treated with S1P (10 μM, 1h). (C) Western blot and (D) quantification of phosphoproteins (p) in Map3k1 TG keratinocytes infected with Ad-GFP or Ad-Cre, with or without MAP2K4/7 inhibitor (BSJ-04-122, BSJ) or JNK inhibitor (SP600125, SP). (E) Quantification of p -PXN (Y) in S1P-treated HEK293 cells (Control, Ctl), cells expressing kinase dead MAP3K1 (KM), or cells with siRNA-mediated MAP3K1 knockdown (siRNA), and in Ad-Cre infected Map3k1TG keratinocytes treated with MAP2K4/7, JNK, or ROCK (Y27632, Y) inhibitors. (F) p -PXN (Y) in S1P-treated wild-type cells with or without ROCK inhibitor (Y) and in Rhoa Δ/Δ ;Rock1 Δ/Δ cells ( R Δ/Δ ;R Δ/Δ ) keratinocytes. (G) Quantification of CN03 (RhoA activator)-induced phosphoproteins in HEK293 cells expressing MAP3K1-KM. (H) S1P-stimulated AP-1 reporter activity and (I) Wound-closure assays in keratinocytes under the indicated genetic perturbations and inhibitor treatments. (J) Schematic model of MAP3K1 signaling network. Data represent mean ± SEM. from ≥3 independent biological replicates. ∗∗∗ p < 0.001 in (B and G) (unpaired two-tailed t test); ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 in (D-F and H-I) (one-way ANOVA followed by Dunnett’s post hoc test).

Article Snippet: Wild type C57BL/6J strain , The Jackson Laboratory , RRID:IMSR_JAX:000664.

Techniques: Western Blot, Infection, Control, Expressing, Knockdown, Activity Assay, Two Tailed Test

Journal: iScience

Article Title: MAP3K1 integrates RhoA/ROCK signaling to regulate epithelial morphogenesis

doi: 10.1016/j.isci.2026.115853

Figure Lengend Snippet: G protein subunits and RhoGEFs connect S1P signaling to RhoA and MAP3K1 activation (A) Schematic model depicting candidate molecular links between S1P receptors, MAP3K1, and RhoA. (B and D) Western blot analyses and (C and E) quantification of phosphoproteins in S1P-treated HEK293 cells in the presence or absence of Gβ inhibitor (Gallein) or siRNAs targeting ARHGEF1 and ARHGEF5 (SC, scrambled control). (F) AP-1 reporter activity in S1P-treated HEK293 cells or MAP3K1-wild type (MAP3K1-WT) adenovirus infected cells with or without pathway inhibitors or ARHGEF knockdown. Data represent mean ± SEM. from ≥3 independent experiments. ∗∗∗ p < 0.001 in (C) (unpaired two-tailed t test); ∗∗ p < 0.01 and ∗∗∗ p < 0.001 in (E and F) (one-way ANOVA followed by Dunnett’s post hoc test).

Article Snippet: Wild type C57BL/6J strain , The Jackson Laboratory , RRID:IMSR_JAX:000664.

Techniques: Activation Assay, Western Blot, Control, Activity Assay, Infection, Knockdown, Two Tailed Test

Journal: iScience

Article Title: MAP3K1 integrates RhoA/ROCK signaling to regulate epithelial morphogenesis

doi: 10.1016/j.isci.2026.115853

Figure Lengend Snippet: G×G and G×E interactions converge on cytoskeleton reorganization (A) Gross eye images at birth (P0) and H&E-stained sections of wild type and Rhoa Δ/Δ ;Rock1 Δ/Δ embryonic eyes at E14.5-E16.5. Red arrows indicate open eye at birth (EOB) defects and the eyelid leading edge. (B) Phalloidin staining and (C) quantification of actin filaments (green) in wild type and Rhoa Δ/Δ ;Rock1 Δ/Δ ( R Δ/Δ ;R Δ/Δ ) eyelid epithelium at E15.5. Left: low magnification; right: enlargements of boxed areas. White arrowheads in (B) mark enriched F-actin at the epithelial leading edge. Dashed lines denote the basement membrane. (D) Phalloidin staining (red) and (E) quantification of F-actin in Rhoa flox/flox ;Rock1 flox/flox keratinocytes infected with Ad-GFP or Ad-GFP-Cre; only GFP positive cells were analyzed. Nuclei are counterstained with Hoechst (blue). Quantification of F-actin intensity in (F) TCDD-treated wild type and Rock1 Δ/Δ pups and (G) Ad-GFP (wild type) and Ad-GFP-Cre infected Rock1 flox/flox ( Rock1 Δ/Δ ) keratinocytes with or without TCDD treatment. Images are representative of at least three mice/genotypes or experimental replicates. CO, cornea; LE, lens; EL, eyelid; RE, retina. Data represent mean ± SEM. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 in (C, E-F) (unpaired two-tailed t test), ∗∗ p < 0.01 in (G) (one-way ANOVA followed by Dunnett’s post hoc test). Scale bars, 500 μm and 200 μm (A) and 50 μm (B and D).

Article Snippet: Wild type C57BL/6J strain , The Jackson Laboratory , RRID:IMSR_JAX:000664.

Techniques: Staining, Membrane, Infection, Two Tailed Test

Journal: iScience

Article Title: MAP3K1 integrates RhoA/ROCK signaling to regulate epithelial morphogenesis

doi: 10.1016/j.isci.2026.115853

Figure Lengend Snippet: Genetic and environmental interactions regulate epithelial differentiation (A) Cell proliferation in wild type and Rhoa Δ/Δ ;Rock1 Δ/Δ ( R Δ/Δ ;R Δ/Δ ) eyelid at E15.5 assessed by EdU labeling. (B) Immunostaining and (C) quantification of Krt1 (green) as a marker of terminal epidermal differentiation in E15.5 eyelids of the indicated genotypes with or without TCDD exposure. Left: low magnification, right: enlarged boxed areas. Dashed lines mark the basement membrane. Nuclei are counterstained with Hoechst (blue). Images represent at least three embryos/genotypes or independent experiments. EL, eyelid. (D and E) RT-qPCR quantification of Krt1 (D) and Krt10 (E) mRNA in Ad-GFP-infected (wild type), Ad-Cre-infected Rock1 flox/flox ( Rock1 Δ/Δ ) and Rhoa flox/flox ; Rock1 flox/flox ( Rhoa Δ/Δ ; Rock1 Δ/Δ ) keratinocytes with or without TCDD treatment. Data represent mean ± SEM. ∗∗ p < 0.01 in (A) (unpaired two-tailed t test) and ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 in (C-E) (one-way ANOVA followed by Dunnett’s post hoc test). Scale bars, 50 μm.

Article Snippet: Wild type C57BL/6J strain , The Jackson Laboratory , RRID:IMSR_JAX:000664.

Techniques: Labeling, Immunostaining, Marker, Membrane, Quantitative RT-PCR, Infection, Two Tailed Test

Journal: Molecular Therapy Advances

Article Title: Swine reporter model for preclinical evaluation and characterization of gene delivery vectors

doi: 10.1016/j.omta.2026.201729

Figure Lengend Snippet: Serum chemistries and blood counts of LNP-injected piglets (A) LNP-mRNA delivery study design schematic. Three 20-day-old SRM-1 piglets were injected with 0.4 mg kg −1 of LNP-mRNA carrying Cre and tissues were harvested after 14 days (B) LNP formulation schematic of mRNA using microfluidic mixing. (C) Absolute values of serum ALT, AST, creatinine, creatinine kinase, and albumin as well as blood cell counts (white blood cells, lymphocytes, neutrophils, red blood cells) in three LNP-injected SRM-1 pigs (purple), one mock-injected SRM-1 pig (gray), and 2 non-injected wild type sibling piglets (black). The red dashed lines are the high and low references ranges supplied by the diagnostic laboratory. One blood sample per piglet was taken at each time point.

Article Snippet: To determine the in vivo stability and delivery efficiency of LNPs with the same formulation used for the systemic pig study, wild-type BALB/cJ mice (male and female, Jackson Laboratory, stock #000651), aged 10–16 weeks and weighing 18–25 g were used for firefly luciferase mRNA delivery experiments.

Techniques: Injection, Formulation, Diagnostic Assay

Journal: Neural Regeneration Research

Article Title: Short-lived Niemann-Pick type C mice with accelerated brain aging as a novel model for Alzheimer’s disease research

doi: 10.4103/NRR.NRR-D-24-01190

Figure Lengend Snippet: Minor Feasibility assessments to characterize motor function loss and ataxia in the NPC1 mut mouse model (both sexes) before the onset of neurological impairment. (A) Mass and (B) survival of WT and NPC1mut mice. No WT animals died during the observation period of 20 weeks, and the median humane survival of NPC1 mut mice was 15 weeks (105 days). n = 9 for all groups for the survival assay. n = 22 for NPC1 mut mice and n = 14 for WT mice for the weight loss study. Data in A represent mean + SD analyzed by mixed-effects model (REML) followed by Šídák’s multiple comparisons test to account for animals that died before week 15, * P < 0.05. (C) Mice were observed daily for the appearance of visible tremors. The median age at which NPC1 mut mice develop tremors was eight weeks. (D–G) Coordination testing of NPC1 mut mice at 13 weeks of age using Rotarod testing at 15 revolutions per minute (D), and a battery of tests (E–G) developed specifically to monitor NPC1-related motor dysfunction as previously described. n = 11 for all groups. (D–G) Data represents mean ± SD and analyzed by unpaired two-tail t -test, * P < 0.05, **** P < 0.0001. NPC: Niemann-Pick type C disease; WT: wild-type.

Article Snippet: Wild-type (WT; C57BL/6) and Npc1 tm(I1061T)Dso , mice (hereafter referred to as NPC1 mut , Strain# 027704, The Jackson Laboratory, Bar Harbor, ME, USA) (Praggastis et al., 2015) were bred and housed under a protocol approved by the VCU Institutional Animal Care and Use Committee (AD10000977, approved December 14, 2022), which has received accreditation from the Association for Assessment and Accreditation of Laboratory Animal Care.

Techniques: Clonogenic Cell Survival Assay, Battery

Journal: Neural Regeneration Research

Article Title: Short-lived Niemann-Pick type C mice with accelerated brain aging as a novel model for Alzheimer’s disease research

doi: 10.4103/NRR.NRR-D-24-01190

Figure Lengend Snippet: Sex-dependent gene expression changes in NPC1 mut mice. (A) The heatmap depicting the similarities and differences in differential expression of significantly altered genes in frontal cortex brain samples across NPC1 mut mice and age- and gender-matched WT controls. (B, C) Venn diagrams depict the commonality analysis results on DEGs from female and male NPC1 mut mice relative to age and gender-matched WT controls. (B) Among the 714 downregulated genes from female samples and 75 from the male mice, 39 genes are common. (C) Similarly, 51 genes are commonly upregulated between female and male mice samples. Additional Table 5 contains the complete list of DEGs, normalized reads, P values, and the common gene list. DEGs: Differentially expressed genes; NPC: Niemann-Pick type C disease; WT: wild-type.

Article Snippet: Wild-type (WT; C57BL/6) and Npc1 tm(I1061T)Dso , mice (hereafter referred to as NPC1 mut , Strain# 027704, The Jackson Laboratory, Bar Harbor, ME, USA) (Praggastis et al., 2015) were bred and housed under a protocol approved by the VCU Institutional Animal Care and Use Committee (AD10000977, approved December 14, 2022), which has received accreditation from the Association for Assessment and Accreditation of Laboratory Animal Care.

Techniques: Gene Expression, Quantitative Proteomics

Journal: Neural Regeneration Research

Article Title: Short-lived Niemann-Pick type C mice with accelerated brain aging as a novel model for Alzheimer’s disease research

doi: 10.4103/NRR.NRR-D-24-01190

Figure Lengend Snippet: Relative transcriptomic age (tAge) and APP/PS1 mice mortality. Relative transcriptomic ages (tAges) and expected lifespans of control and APP/PS1 mouse brain samples were estimated with the multi-tissue clock, (A) chronological tAge, (B) expected lifespan (mon), in comparison to the corresponding WT control mice. WT: Wild-type.

Article Snippet: Wild-type (WT; C57BL/6) and Npc1 tm(I1061T)Dso , mice (hereafter referred to as NPC1 mut , Strain# 027704, The Jackson Laboratory, Bar Harbor, ME, USA) (Praggastis et al., 2015) were bred and housed under a protocol approved by the VCU Institutional Animal Care and Use Committee (AD10000977, approved December 14, 2022), which has received accreditation from the Association for Assessment and Accreditation of Laboratory Animal Care.

Techniques: Control, Comparison

Journal: Neural Regeneration Research

Article Title: Short-lived Niemann-Pick type C mice with accelerated brain aging as a novel model for Alzheimer’s disease research

doi: 10.4103/NRR.NRR-D-24-01190

Figure Lengend Snippet: Immunohistochemical analysis of microglia and astrocytes in the Hippocampus of male and female mice of NPC1 mut mice. Confocal images of the Hippocampus from (A) female and (B) male mice in two groups: control (CTRL) and NPC1 mut mice. Astrocytes are labeled in red (GFAP), microglia in green (Iba1), and nuclei in blue (DAPI). An increase in microglial density is observed in the NPC1 group compared to the CTRL group in both genders. Quantitative comparison of microglial count (Iba1 + cells) in female (C) and male mice (D) across the two groups, showing a significant increase in microglial numbers in the NPC1 group compared to the CTRL group (two-tailed t -test; P < 0.05). DAPI: 4’,6-Diamidino-2-phenylindole; GFAP: glial fibrillary acidic protein; Iba-1: ionized calcium-binding adapter molecule 1; NPC: Niemann-Pick type C disease; WT: wild-type.

Article Snippet: Wild-type (WT; C57BL/6) and Npc1 tm(I1061T)Dso , mice (hereafter referred to as NPC1 mut , Strain# 027704, The Jackson Laboratory, Bar Harbor, ME, USA) (Praggastis et al., 2015) were bred and housed under a protocol approved by the VCU Institutional Animal Care and Use Committee (AD10000977, approved December 14, 2022), which has received accreditation from the Association for Assessment and Accreditation of Laboratory Animal Care.

Techniques: Immunohistochemical staining, Control, Labeling, Comparison, Two Tailed Test, Binding Assay