Journal: Nature Communications

Article Title: Hypoxia induces HIF1α-dependent epigenetic vulnerability in triple negative breast cancer to confer immune effector dysfunction and resistance to anti-PD-1 immunotherapy

doi: 10.1038/s41467-022-31764-9

Figure Lengend Snippet: a Schematic diagram showing the establishment of humanized mice (humice) with human immune system reconstituted in NIKO mice. The presence of human CD45 + cells, NK cells, CD4 + and CD8 + T cells in the mice’s peripheral system was validated by flow cytometry. b Primary LM2 tumor size in humice (control, n = 14; Keytruda, n = 14; ENT, n = 12; PX478, n = 14; ENT + Keytruda, n = 16; PX478 + Keytruda, n = 16) and NIKO mice (control, n = 10; ENT + Keytruda, n = 10; PX478 + Keytruda, n = 10), at Day 21 of treatments. c Lung metastasis of humice (control, n = 6; Keytruda, n = 6; ENT, n = 6; PX478, n = 6; ENT + Keytruda, n = 7; PX478 + Keytruda, n = 7) and NIKO mice (control, n = 5; ENT + Keytruda, n = 5; PX478 + Keytruda, n = 5) bearing LM2 tumors at Day 35 assessed by bioluminescence (BLI) measurement. d Representative bioluminescence (BLI) images showing the lung metastasis of humice and NIKO mice. e Flow cytometric analysis of LM2 tumors harvested from humanized mice. IFNγ, TNFα, and granzyme B expression was examined in tumor-infiltrating human CD8 + T cells and NK cells. N = 5 for each group. f Flow cytometry analysis of LM2 tumors harvested from humanized mice. Expressions of human PD-L1 and PD-L2 were examined in total living cells dissociated from LM2 tumors. N = 5 for each group. Quantification data of flow cytometry ( e , f ) are presented as a box and whiskers, with median values and whiskers of minimum and maximum values. Data for b and c were presented as mean ± SD . P values were determined by one-way ( e , f ) or two-way ( b , c ) ANOVA with Turkey’s test. Source data are provided as a source data file.

Article Snippet: The following antibodies were used for staining, anti-activated pimonidazole FITC antibody (Hypoxyprobe, CAT# HP2-200kit, dilution 1:200), anti-mouse HIF1α APC antibody (R&D Systems, CAT# IC1935A, dilution 1:50), anti-mouse CD3 BV421 antibody (BD Biosciences, CAT# 564008, dilution 1:100), anti-mouse CD45 Percp-Vio700 antibody (Miltenyi Biotec, CAT# 130-110-663, dilution 1:100) anti-mouse CD8 APC-Vio770 antibody (Miltenyi Biotec, CAT# 130-120-737, dilution 1:100), anti-mouse Nkp46 APC antibody (Miltenyi Biotec, CAT# 130-112-202, dilution 1:100), anti-mouse CD4 BV650 antibody (Biolegend, CAT# 563747, dilution 1:100), anti-mouse TIM-3 BV711 antibody (Biolegend, CAT# 119727, dilution 1:100), anti-mouse PD-1 PE-Vio770 (Miltenyi Biotec, CAT# 130-120-391, dilution 1:100), anti-mouse IFNγ PE (Miltenyi Biotec, CAT# 130-117-352, dilution 1:100), anti-mouse TNFα BV711 (BD Biosciences, CAT# 563944, dilution 1:100), anti-mouse/human granzyme B FITC (Miltenyi Biotec, Cat#130-118-430, dilution 1:100), anti-mouse PD-L1 BV786 antibody (BD Biosciences, CAT# 741014, dilution 1:100), anti-mouse PD-L2 FITC antibody (Miltenyi Biotec, Cat# 130-102-222, dilution 1:100), anti-human CD45 FITC antibody (BD Biosciences, CAT# 304006, dilution 1:100), anti-human CD3 PE antibody (Biolegend, CAT# 300308, dilution 1:100) anti-human CD8 APC-Cy7 antibody (BD Biosciences, CAT# 557834, dilution 1:100), anti-human CD56 BV711 antibody (Biolegend, CAT# 318336, dilution 1:100), anti-human CD4 APC antibody (Biolegend, CAT# 300514, dilution 1:100), anti-human IFNγ BV785 (Biolegend, CAT# 502542, dilution 1:100), anti-human TNFα BV650 (Biolegend, CAT# 502398, dilution 1:100), anti-human Granzyme B BV421 (BD Biosciences, Cat# 563389, dilution 1:100), anti-human PD-L1 PE-Cy7 antibody (Biolegend, CAT# 374506, dilution 1:100), anti-human PD-L2 PE antibody (Miltenyi Biotec, CAT# 130-098-530, dilution 1:100).

Techniques: Flow Cytometry, Control, Expressing

Journal: bioRxiv

Article Title: A class act: HDAC1- Malat1 regulates MDSC apoptosis and cell cycling to decrease suppression of T cells

doi: 10.64898/2026.03.23.713743

Figure Lengend Snippet: (A) UMAP visualization of major cell clusters identified from single cell RNA sequencing (scRNA-seq) of NT2.5-LM breast-to-lung metastatic tumors after 3 weeks of treatment with: vehicle (V), Entinostat (E), anti-CTLA-4 + anti-PD-1 immune checkpoint inhibitors (PC), and Entinostat + anti-CTLA-4 + anti-PD-1 combination (EPC). Subclusters of MDSCs identified: G-MDSCs and M-MDSCs. (B) Iterative logistic regression analyses conducted on the major MDSC cluster comparing the Vehicle (V) and Entinostat + ICIs combination treatment (EPC). (C) Expression of Malat1 / MALAT1 in G- and M-MDSCs isolated from lung metastases of NT2.5-LM mice treated with vehicle vs. Entinostat for 3 weeks (left, n=2 from 5 pooled mice per treatment group), J774M murine MDSC-like cell line treated with Entinostat for 24 hours (middle, n=3), and human PBMC-derived MDSCs (hMDSCs) treated with Entinostat for 24 hours (right, n=3). (D) Median Fluorescence Intensity expression of Malat1 in CD45 + CD11b + MHC-II - F4/80 - Ly6G - Ly6C hi cells from lung metastases in 4T1 mice, treated with vehicle (V), Entinostat (E), and Entinostat + ICIs (EPC) for 3 weeks. (E) Expression of Malat1 in J774M cell line treated with various concentrations of various HDAC inhibitors, with corresponding targeted class and specific HDACs. One-way ANOVA for (C, E: all statistically significant with p<0.05, unless indicated as non-significant (ns)), Kruskal-Wallis test with Dunn’s correction for (D). * p<0.05, *** p< 0.001, **** p<0.0001. ENT = Entinostat; PAN = Panobinostat; BEL = Belinostat; VOR = Vorinostat; TSA = Trichostatin A; DOM = Domatinostat; CHL = Chlopynostat; PYR = Pyroxamide; SCA = Santacruzamate A; RGF = RGFP966; TMP = TMP269; SIS = SIS17.

Article Snippet: Immune checkpoint inhibitors (ICIs) used in this study: anti-CTLA-4 (BioXCell cat. #BE0131) and anti-PD-1 (BioXCell cat. #BE0146).

Techniques: Single Cell, RNA Sequencing, Expressing, Isolation, Derivative Assay, Fluorescence

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A ) Mice were inoculated with 3×10 6 BPPNM cells IP and dosed with 20 μg of STING or Scr conjugate delivered by LNP IP at 10, 13, 16, and 19 days after inoculation. Omental tumor was collected on day 20 for analysis by flow cytometry. Groups included N = 5 mice. ( B ) Cell populations in tumor after treatment, displaying percentage of CD45 + cells made up by T cells (CD3 + ), CD8 + T cells, CD4 + T cells, NK cells (CD3 - NK1.1 + ), B cells (CD19 + ), Macrophages (F4/80 + ), DCs (CD11c + MHCII + ), and MDSCs (CD11b + Gr-1 hi ). Percentage of CD45 - cells made up by BPPNM cancer cells (GFP + ) are also displayed. ( C-D ) Polarization of Macrophages (F4/80 + ), showing MFI and representative distributions of ( C ) CD86 and ( D ) CD206. ( E ) Activation of DCs (CD11c + MHCII + ), showing MFI and representative distributions of CD86. ( F ) Expression of PD-1 on CD8 + T cells, showing percentage of cells that are PD-1 + and representative distributions of PD-1. P values computed with a one-way ANOVA followed by Tukey’s post-hoc test are displayed above each figure. All flow data is represented as mean ± SD. ( G ) Mice were inoculated with 3×10 6 BPPNM cells IP and dosed with 20 μg of STING or Scr conjugate delivered by LNP IP at 10, 13, 16, 19, and 22 days after inoculation. A subset of groups were additionally treated with 100 μg αPD-1 antibody at 11 and 17 days after inoculation. Groups included N = 6 (PBS, STING LNP, Scr LNP), N = 5 (STING LNP + αPD-1), or N = 4 (PBS + αPD-1, Scr LNP + αPD-1) mice. ( H ) Survival plot with P values determined by log(rank) (Mantel–Cox) test. ( I-K ) Tumor burden measured by IVIS, displaying ( I ) geometric mean ± SD bioluminescent intensity over the treatment period, ( J ) an image of the mouse with the median bioluminescent intensity from each group at the measurement following the end of treatment (day 24), and ( K ) individual mouse bioluminescent intensity values.

Article Snippet: In groups specified in figure captions, mice were also administered a 100 μg dose of anti-mouse PD-1 antibody (Bio X Cell #BE0273 InVivoMAb, Clone 29F.1A12) in 200 μL of 1× PBS on days 11 and 17 after tumor inoculation.

Techniques: Flow Cytometry, Activation Assay, Expressing