Journal: Future Science OA

Article Title: Phosphoglucose isomerase directs the inflammatory response, calcium influx and fibroblast migration in keloids

doi: 10.1080/20565623.2026.2615968

Figure Lengend Snippet: PGI expression was increased in hyperfibrotic regions in keloid tissues. (A) Immunohistochemical staining for PGI in normal skin and keloids (original magnification 40). Right panels are high-power views of the tissues (original magnification 200). The black rectangles in the left panels indicate the areas enlarged at high magnification, respectively. The PGI intensity is shown (B). (C) Sections of keloid tissues and normal skin were deparaffinized and further incubated with antibodies against PGI and vimentin. The expression of PGI (green) and vimentin (red) was localized in keloid tissue and normal skin (original magnification 100). Right panels are high-power views of tissues, identifying colocalization of PGI (green) and vimentin (red) in keloid and normal skin (original magnification 200). The white rectangles in the left panels show the enlarged areas at high magnification, respectively. The number of PGI-positive, vimentin-expressing cells was quantified from five randomly selected high-power fields. The percentage of these cells normalized to total cells (DAPI-stained cells) in the hypercellular areas is shown (D). Data are presented as the mean ± SD. (** p < 0.01) from representative data of 3 independent experiments.

Article Snippet: Sections were applied with 1:100-diluted rabbit anti-human PGI (#57893, Cell Signaling) and 1:500-diluted mouse anti-human vimentin primary antibodies (#5741, Cell Signaling) at 4 °C overnight followed by washing and incubated with secondary antibodies conjugated with 1:500-diluted goat anti-mouse Alexa Fluor TM 488 (A11001, Invitrogen) and goat anti-rabbit Alexa Fluor TM 568 (A11011, Invitrogen).

Techniques: Expressing, Immunohistochemical staining, Staining, Incubation

Journal: NPJ Breast Cancer

Article Title: Agonistic CD40 elicits CD8+ T-cell-dependent primary responses and CD4+ T-cell-dependent long-term immunity in breast cancer

doi: 10.1038/s41523-025-00889-7

Figure Lengend Snippet: A Proportions of CD3+, CD8 +, and CD4 + T cells by flow cytometry in control and aCD40 treated Brpkp110 tumors on day 13 post treatment ( n = 10). B FoxP3 + CD4+ regulatory T cells on day 7 post-treatment ( n = 7). C Proportions of Granzyme B + T cells in subpopulations by flow cytometry in control and aCD40 treated Brpkp110 tumors on day 7 post implantation ( n = 10). D Images of immunofluorescent staining for CD8 (red) and nuclei (blue) and CD8 staining quantification in untreated (control) and aCD40 treated Brpkp110 tumors on day 7 post treatment ( n = 4–6). E Quantification of CD8 immunofluorescent staining in outer, middle, and inner thirds of control and aCD40 treated tumors on day 7 post treatment ( n = 5–6). F After 7 days of treatment with aCD40, Brpkp110 tumors were minced and cultured ex vivo. Supernatant was collected and pooled for each treatment group after 48 h and cytokines were measured ( n = 2, with 3 tumors pooled per group). G T cell activation and proliferation markers measured by flow cytometry in CD4+ and CD8 + T cell populations of control and aCD40 treated Brpkp110 TDLN on day 7 post-treatment ( n = 10). H Growth curves of control and aCD40 treated Brpkp110 tumors implanted into WT hosts with or without T cell depletions ( n = 17–20). I Tumor growth curve of aCD40 (treatment on Day 8) +/− FTY720 (treatment started on Day 7) treated Brpkp110 tumors ( n = 14–16). Data: ( A – C , D (right), E – G ) median, ( H , I ) mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Tissue sections were blocked with 5% donkey serum diluted in PBS with 0.3% Triton X-100 and stained with rabbit anti-mouse primary CD8 (clone: D4W2Z, Cell Signaling Technology 98941, 1:100) followed by donkey anti-rabbit Alexa Fluor 594 (Thermo Scientific A-21207, 1:250) secondary antibody.

Techniques: Flow Cytometry, Control, Staining, Cell Culture, Ex Vivo, Activation Assay

Journal: NPJ Breast Cancer

Article Title: Agonistic CD40 elicits CD8+ T-cell-dependent primary responses and CD4+ T-cell-dependent long-term immunity in breast cancer

doi: 10.1038/s41523-025-00889-7

Figure Lengend Snippet: A Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 26 post implantation (right) of Brpkp110 tumors. Indicated treatments initiated on day 7 post implantation ( n = 18–20). B Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 26 post implantation (right) of E0771 tumors. Indicated treatments initiated on day 8 post implantation ( n = 12–15). C Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 31 post implantation (right) of AT3 tumors. Indicated treatments initiated on day 9 post implantation ( n = 14). D Tumor growth curves (left) and tumor volume changes compared to pretreatment on day 34 post implantation (right) of EpH4 1424 tumors. Indicated treatments initiated on day 5 post implantation ( n = 14–16). E Brpkp110 tumor growth curves (left) and volume changes compared to pretreatment (right) on day 27 post implantation in control and aCD40+ICB treated hosts with and without CD8 + T cell depletions. Indicated treatments initiated on day 7 post implantation ( n = 12–15). F Brpkp110 tumor growth curves (left) and volume changes compared to pretreatment (right) on day 25 post implantation in control and aCD40+ICB treated hosts with and without CD4 + T cell depletions. Indicated treatments initiated on day 8 post implantation ( n = 14–18). G Brpkp110 tumor growth curves (left) and volume changes compared to pretreatment (right) on day 27 post implantation in control and aCD40+ICB treated hosts with and without CD4+ and CD8 + T cell depletions. Indicated treatments initiated on day 7 post implantation ( n = 12–18). H Tumor growth curves (left) and volume changes compared to pretreatment (right) on day 26 post Brpkp110 tumor implantation into WT and BATF3 KO hosts. Indicated treatments initiated on day 7 post implantation ( n = 14–18). Data: ( A – H left) mean ± SEM, ( A – H right) each column represents individual tumor. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Tissue sections were blocked with 5% donkey serum diluted in PBS with 0.3% Triton X-100 and stained with rabbit anti-mouse primary CD8 (clone: D4W2Z, Cell Signaling Technology 98941, 1:100) followed by donkey anti-rabbit Alexa Fluor 594 (Thermo Scientific A-21207, 1:250) secondary antibody.

Techniques: Control, Tumor Implantation

Journal: NPJ Breast Cancer

Article Title: Agonistic CD40 elicits CD8+ T-cell-dependent primary responses and CD4+ T-cell-dependent long-term immunity in breast cancer

doi: 10.1038/s41523-025-00889-7

Figure Lengend Snippet: A Proportions of circulating effector memory (CD44 + CD62L-), central memory (CD44 + CD62L+), and naïve (CD44-CD62L-) CD4+ (left) and CD8+ (right) in blood, 3 months post treatment induced tumor clearance ( n = 5–6, data representative of 2 experiments with similar results). B Secondary Brpkp110 tumor rechallenge of naïve and previously Brpkp110 tumor-bearing mice cured after aCD40 + ICB, at least 2 months post primary tumor clearance ( n = 12–14, data representative of 3 experiments with similar results). C Control and rechallenge tumor growth in T cell sufficient ( n = 6–12) and T cell depleted hosts ( n = 12–14, data representative of 2 experiments with similar results). D Brpkp110 tumor growth curves in intra-tumoral (IT) vehicle (control) and IT aCD40 treated hosts. aCD40 administered tumors denoted as aCD40 IT and contralateral untreated tumors denoted as CD40 IT Distant ( n = 9–12, data representative of 2 experiments with similar results). E Brpkp110 tumor growth curves in intra-tumoral (IT) vehicle (control) and IT or intraperitoneal (IP) aCD40 or ICB received hosts ( n = 4–9, data representative of 2 experiments with similar results). Data: ( A ) median, ( C – E ) mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Tissue sections were blocked with 5% donkey serum diluted in PBS with 0.3% Triton X-100 and stained with rabbit anti-mouse primary CD8 (clone: D4W2Z, Cell Signaling Technology 98941, 1:100) followed by donkey anti-rabbit Alexa Fluor 594 (Thermo Scientific A-21207, 1:250) secondary antibody.

Techniques: Control

Journal: bioRxiv

Article Title: Wnt/β-catenin signaling promotes zebrafish osteoblast dedifferentiation by wnt10a -mediated inhibition of NF-κB

doi: 10.64898/2025.12.29.696582

Figure Lengend Snippet: A) RNASeq (left) and RT-qPCR (right) reveal upregulation of the NF-κB signaling target genes nfkbiaa and nfkbiab in osteoblasts sorted from bglap:GFP fish heterozygous for the wnt10a mutation relative to their wild-type siblings at 1 dpa. nE (qPCR biological replicates) = 3, nA = 10 per replicate. ΔΔCt values are normalized to the mean of the wildtype at 1 dpa. Error bars, Mean ± SEM. Two tailed Student’s t-test. B) Osteoblast dedifferentiation, as measured by bglap downregulation in segment -1 revealed by HCR in situ hybridization, is inhibited in fish treated with the Wnt inhibitor IWR-1, while IP injection of the NF-κB inhibitor Bay-11 slightly but significantly enhances dedifferentiation. Treatment with both inhibitors yields results similar to those of Bay-11 alone. nE = 3 (except for 2 for IWR-1), nA = 18 total per group (12 for IWR-1), nR = 33 (DMSO), 24 (IWR-1), 37 (Bay-11), 33 (both). C) Overexpression of wnt10a using hs:wnt10a fish is sufficient to cause downregulation of bglapl detected by HCR in situ hybridization in non-injured fins, relative to heat-shocked wild-type fish. HCR signals were quantified in a bony segment (“B”) that is located at the same proximal-distal position as “segment -1” in amputated fins. nE = 2, nA = 12 total per group, nR = 22 total per group. Dashed line, joints. Scale bar, 100 µm. D) Immunofluorescence on cryosections of hs:wnt10a transgenic hearts reveals increased embryonic myosin heavy chain (embMHC) expression in Myl7+ cardiomyocytes at the wound border at 7 days post injury (dpi). Plots show the ventricular area covered by anti-embMHC staining relative to the 150 µm wound border zone area occupied by Myl7+ myocardium. nE = 2, nA = 13 wild-type, 11 hs:wnt10a . Scale bar, 100 µm. (B, C, D) Error bars, mean ± 95% CI. Two tailed Student’s t-test.

Article Snippet: Primary antibodies against embMHC Mouse monoclonal (MYH7 DSHB, N2.261, RRID:AB_531790), and Myl7 Rabbit polyclonal (GeneTex, GTX128346, RRID:AB_2885759) were diluted in PEMTx/normal goad serum and applied overnight at 4 °C.

Techniques: Quantitative RT-PCR, Mutagenesis, Two Tailed Test, In Situ Hybridization, Injection, Over Expression, Immunofluorescence, Transgenic Assay, Expressing, Staining

Journal: Molecular Cancer Therapeutics

Article Title: The Anti-FRα Antibody–Drug Conjugate Luveltamab Tazevibulin Demonstrates Efficacy in Non–Small Cell Lung Cancer Preclinical Models and Induces Immunogenic Cell Death

doi: 10.1158/1535-7163.MCT-24-0649

Figure Lengend Snippet: Luveltamab tazevibulin induces hallmarks of ICD and can activate monocytes in tumor cell–PBMC cell co-cultures in vitro . Fold change in ( A ) HMGB1 release, ( B ) ATP release, and ( C ) calreticulin expression in KB (FRα + ) and A549 (FRα − ) tumor cell cultures with medium only or 50 nmol/L isotype control ADC, SC209, or luveltamab tazevibulin ( n = 3 replicates per group). Supernatants and cells were analyzed after 72 hours of incubation. Fold change in comparison with the medium-only group is depicted. For calreticulin expression, representative histograms are shown to the right. Dose–response curves of the proportion of ( D ) CD86 + cells or ( E ) CellTrace + cells among CD14 + immune cells in tumor cell–PBMC co-cultures incubated with isotype control ADC, SC209, or luveltamab tazevibulin for 48 hours at the indicated concentrations ( n = 2 replicates at each concentration level). Dose–response curves from two separate donors are shown. Fold change in ( F ) HMGB1 release, ( G ) ATP release, and ( H ) calreticulin expression in MC38-hFRα and MC38 wild-type tumor cell cultures with medium only or 50 nmol/L SC209 or luveltamab tazevibulin ( n = 3 replicates per group). Supernatants and cells were analyzed after 48 hours of incubation. Fold change in comparison with the medium-only group is depicted. For calreticulin expression, representative histograms are shown to the right. I, Dose–response curves of the proportion of CD86 + cells among CD14 + immune cells in human PBMC–tumor cell co-cultures incubated with isotype control ADC, SC209, or luveltamab tazevibulin for 48 hours at the indicated concentrations ( n = 2 replicates at each concentration level). Data are presented as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001 estimated by a one-way ANOVA with the Dunnett multiple comparison test.

Article Snippet: ICD markers following the treatment of MC38-hFRα and MC38 wild-type cells were performed in a similar manner as above, with minor differences: cells were treated for 48 hours before analysis of ICD markers; calreticulin was detected using a rabbit anti–mouse calreticulin primary antibody (#12238, Cell Signaling Technology) and a goat anti–rabbit IgG Alexa Fluor 647 secondary antibody (Invitrogen); and HMGB1 detection was performed using Ultra TMB and TMB BlueSTOP Solution (Seracare) and read at 650 nm.

Techniques: In Vitro, Expressing, Control, Incubation, Comparison, Concentration Assay