Journal: Molecular Therapy Oncology

Article Title: Recombinant CALR polarizes and activates macrophages in tumors

doi: 10.1016/j.omton.2025.201121

Figure Lengend Snippet: Recombinant CALR reduces tumor growth and increases M1 macrophages (A) To measure the effect on tumor growth and immune cell activation, bacterial lysate was injected into tumor-bearing mice. Tumors were formed by subcutaneously injecting CT26 murine colon carcinoma cells into the flank of BALB/c mice. After 2 weeks, the mice were injected intratumorally with saline, bacterial control lysate, or lysate from CALR-expressing bacteria. One set of mice received injections at days 0, 3, and 6; were monitored for tumor growth; and their tumors were harvested at day 9 for analysis of immune cells. A second set of mice received only one injection at day 0, and tumors were harvested at day 3 for analysis. (B) Intratumoral injection of CALR lysate decreased tumor growth compared to saline controls ( p = 0.0089). Bacterial control lysate also reduced tumor growth compared to controls ( p = 0.0204). Volumes are reported relative to those on day 0. (C–F) On day 3, recombinant CALR did not affect (C) the number of leukocytes, (D) the number of M1 macrophages (per 10,000 cells analyzed), or the number of M1 macrophages expressing either (E) CD80 or (F) CD86 (per 10,000 cells analyzed) in tumors. (G) On day 9, injection with CALR lysate significantly increased the number of leukocytes in tumors compared to saline controls ( p = 0.0042). (H) On day 9, CALR lysate also significantly increased the number of M1 macrophages in tumors compared to bacterial controls ( p = 0.0480) and saline ( p = 0.0061). (I) CALR lysate significantly increased the number of M1 macrophages expressing CD80 (per 10,000 cells analyzed) compared to saline controls ( p = 0.0063). (J) CALR lysate also increased the number of M1 macrophages expressing CD86 (per 10,000 cells analyzed) compared to bacterial controls ( p = 0.0445) and saline ( p = 0.0077). Data are represented as mean ± SEM. The statistical comparisons in (B) are two-way ANOVA followed by Tukey’s method. The statistical comparisons in (C–J) are ANOVA followed by Tukey’s method. Asterisks indicate significance: ∗ p < 0.05; ∗∗ p < 0.01.

Article Snippet: JAWSII murine dendritic cells and CT26 murine colon carcinoma cells were obtained from ATCC, confirmed by STR profiling by Charles River Research Animal Diagnostic Services, and passaged for fewer than 6 months.

Techniques: Recombinant, Activation Assay, Injection, Saline, Control, Expressing, Bacteria

Journal: Molecular Therapy Oncology

Article Title: Recombinant CALR polarizes and activates macrophages in tumors

doi: 10.1016/j.omton.2025.201121

Figure Lengend Snippet: Recombinant CALR increases helper T cell activity in tumors (A) The extent of T cell infiltration was determined in mice (see ) with CT26 tumors that were intratumorally injected (on days 0, 3, and 6) with saline (PBS), bacterial control lysate (BC), or lysate from CALR-expressing bacteria (CALR). On day 9, injection of CALR lysate significantly increased the number of T cells in tumors (per 10,000 cells analyzed) compared to saline controls ( p = 0.0495). (B) The percentage of helper T cells per 10,000 cells analyzed. (C) On day 9, injection with CALR lysate significantly increased the number of activated helper T cells (per 10,000 cells analyzed) in tumors compared to saline controls ( p = 0.0067). Data are represented as mean ± SEM. The statistical comparisons in (A–C) are ANOVA followed by Dunnett’s test. Asterisks indicate significance: ∗ p < 0.05; ∗∗ p < 0.01.

Article Snippet: JAWSII murine dendritic cells and CT26 murine colon carcinoma cells were obtained from ATCC, confirmed by STR profiling by Charles River Research Animal Diagnostic Services, and passaged for fewer than 6 months.

Techniques: Recombinant, Activity Assay, Injection, Saline, Control, Expressing, Bacteria

Journal: Journal of Human Immunity

Article Title: Complete and partial forms of X-linked MCTS1 deficiency in patients with mycobacterial disease

doi: 10.70962/jhi.20250073

Figure Lengend Snippet: Functional investigation of the new MCTS1 variants. (A) Predicted structure of the MCTS1 protein with the domain of unknown function 1947 (DUF1947) and the pseudouridine synthase and archaeosine transglycosylase (PUA) domains for WT protein, the synthetic LOF variant p. A109D, and the patient variants p.L170* (P1), p.W175* (P2), and predicted p.E60Kfs5* (P4). The numbers annotated for the WT MCTS1 protein correspond to the first and last amino acids of the MCTS1 protein, the DUF1947, and PUA domains. The variants of MCTS1 are shown in red. (B) Western blot of WT and MCTS1-KO HeLa cells after transfection with EV, WT MCTS1 (pWT), or the MCTS1 variants p.E60Kfs5* (pE60Kfs5*), p.L170* (pL170*), p.W175* (pW175*), and p.A109D (pA109D). Details are provided in . Non-transfected HeLa cells (“−”) were used as a negative control. (C) Schematic diagram of the Fluc and lamin B + stuORF reporters used to assess MCTS1 activity. stuORF, synthetic strong Kozak uORF. Modified from . (D) Activity of the MCTS1 variants in the luciferase translation reinitiation assay after the transfection of WT and MCTS1-KO cells with EV, WT MCTS1 or the various MCTS1 variants (p.E60Kfs5* p.L170*, W175*, and p.A109D; see details in ). Bars: Mean and standard deviation of three technical replicates, represented as turquoise dots. This experiment is representative of the three biological replicates. The asterisks indicate the level of significance, as assessed in a one-way ANOVA with Tukey’s test for multiple comparisons and adjustment for multiple testing (*P < 0.05; **P < 0.005). (E) Sanger sequencing confirmation of KI of the MCTS1 W175* variant and silent L174L variants in HEK293T cells. (F) Western blot of HEK293T cells subjected to genome editing to introduce MCTS1 KO in a pooled manner or MCTS1 W175* KI with the indicated single-cell derived clones shown. (G) Quantitation of three independent western blots of the indicated cell lines. Statistical significance was assessed using two-sided Mann–Whitney tests. Source data are available for this figure: .

Article Snippet: WT HeLa cells (Cat# CRM-CCL-2; ATCC, RRID: CVCL_0030) and MCTS1 KO HeLa cells ( ) were cultured in Dulbecco/Vogt modified Eagle’s minimal essential medium (DMEM, Gibco) with 10% fetal-calf serum (FCS) and 100 IU/ml penicillin/streptomycin (Cat# 15140122; Gibco).

Techniques: Functional Assay, Variant Assay, Western Blot, Transfection, Negative Control, Activity Assay, Modification, Luciferase, Standard Deviation, Sequencing, Introduce, Derivative Assay, Clone Assay, Quantitation Assay, MANN-WHITNEY

Journal: iScience

Article Title: Fluorescence tracking Treg movement identifies anti-CCR8 and radiation as a therapeutic combination

doi: 10.1016/j.isci.2025.114572

Figure Lengend Snippet: Combination RT and anti-CCR8 reduces tumor burden and improve survival in distant non-irradiated tumors BALB/c mice were injected with CT26 on both flanks. Tumor-bearing mice were treated with a “pre” treatment strategy. Mice were injected i.p. with anti-CCR8 on days 7, 10, and 14 and treated with 12 Gy RT to the right flank tumor (treated) on day14 post-tumor inoculation. The left tumor (untreated) did not receive RT. Tumor growth and survival were assessed. A total of 8 mice were analyzed per group. Data are represented as mean ± SD. Statistics for survival plots were performed using a Mantel-Cox test between groups. ns = not significant, ∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05.

Article Snippet: The CT26 murine colorectal carcinoma cell line was purchased from ATCC (CRL-2638).

Techniques: Irradiation, Injection

Journal: iScience

Article Title: Fluorescence tracking Treg movement identifies anti-CCR8 and radiation as a therapeutic combination

doi: 10.1016/j.isci.2025.114572

Figure Lengend Snippet: Combination RT and anti-CCR8 reduces tumor burden and improve survival in distant non-irradiated tumors BALB/c mice were injected with CT26 on both flanks. Tumor-bearing mice were treated with a “pre” treatment strategy. Mice were injected i.p. with anti-CCR8 on days 7, 10, and 14 and treated with 12 Gy RT to the right flank tumor (treated) on day14 post-tumor inoculation. The left tumor (untreated) did not receive RT. Tumor growth and survival were assessed. A total of 8 mice were analyzed per group. Data are represented as mean ± SD. Statistics for survival plots were performed using a Mantel-Cox test between groups. ns = not significant, ∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05.

Article Snippet: No additional authentication was performed beyond obtaining MC38, Moc1, and Moc2 from collaborating laboratories that routinely use and maintain these models, and CT26 from ATCC.

Techniques: Irradiation, Injection

Journal: iScience

Article Title: Fluorescence tracking Treg movement identifies anti-CCR8 and radiation as a therapeutic combination

doi: 10.1016/j.isci.2025.114572

Figure Lengend Snippet: Combination RT and anti-CCR8 reduces tumor burden and improve survival in distant non-irradiated tumors BALB/c mice were injected with CT26 on both flanks. Tumor-bearing mice were treated with a “pre” treatment strategy. Mice were injected i.p. with anti-CCR8 on days 7, 10, and 14 and treated with 12 Gy RT to the right flank tumor (treated) on day14 post-tumor inoculation. The left tumor (untreated) did not receive RT. Tumor growth and survival were assessed. A total of 8 mice were analyzed per group. Data are represented as mean ± SD. Statistics for survival plots were performed using a Mantel-Cox test between groups. ns = not significant, ∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05.

Article Snippet: CT26 mouse colorectal carcinoma , ATCC , Cat #: CRL-2638.

Techniques: Irradiation, Injection

Journal: Biogerontology

Article Title: Tumor-derived circulating DNA can induce senescence and SASP activation in mouse embryonic fibroblasts

doi: 10.1007/s10522-026-10400-9

Figure Lengend Snippet: Quantitative analysis of senescence associated gene expression in MEFs following 24 h cfDNA or ctDNA exposure. a p16, b p21, c p53

Article Snippet: Mouse embryonic fibroblasts (ATCC CRL-2991) were cultured at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Gene Expression

Journal: Biogerontology

Article Title: Tumor-derived circulating DNA can induce senescence and SASP activation in mouse embryonic fibroblasts

doi: 10.1007/s10522-026-10400-9

Figure Lengend Snippet: Quantitative analysis of SASP associated gene expression in MEFs following 24 h cfDNA or ctDNA exposure. a IL-1B, b IL-6

Article Snippet: Mouse embryonic fibroblasts (ATCC CRL-2991) were cultured at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Gene Expression

Journal: Biogerontology

Article Title: Tumor-derived circulating DNA can induce senescence and SASP activation in mouse embryonic fibroblasts

doi: 10.1007/s10522-026-10400-9

Figure Lengend Snippet: Senescence-associated β-galactosidase staining of MEFs following 24 h exposure to cfDNA or ctDNA. Images captured under brightfield microscopy using a 10 × objective. a NC, b Sen ( +) /PC, c ctDNA-LD, d ctDNA-HD, e cfDNA-LD, f cfDNA-HD

Article Snippet: Mouse embryonic fibroblasts (ATCC CRL-2991) were cultured at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Staining, Microscopy