Journal: Advanced Science

Article Title: Ir(III) Complexes Convert Cold to Hot Tumors via Ferroptosis/Necroptosis‐Driven Immunogenic Cell Death and Photosensitized CD47 Downregulation

doi: 10.1002/advs.202514256

Figure Lengend Snippet: Ir1‐PDT establishes protective immunity against tumor rechallenge . (a) Schedule of in vivo evaluation. (b) Right‐sided tumor growth curves over time in different treatment groups (n = 5). (c) Left‐sided tumor growth curves over time in different treatment groups (n = 5). (d) Body weight changes in mice during the experimental period (n = 5). (e) Photographic documentation of tumors harvested at endpoint (Day 17, n = 5). (f) Immunohistochemical and hematoxylin and eosin (H&E) staining of right‐sided tumors. Scale bar: 50 µm. (g‐m) Proportion of immune cell subsets in right‐sided tumors. (n and o) Representative immunofluorescence images of left‐sided tumor‐infiltrating (n) CD4 + (red) and (o) CD8 + (green) T cells. Nuclei counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI, blue). Scale bar: 50 µm. Groups: Dark (i), Light (ii), Ir1 (1 mg kg −1 ) + dark (iii), Ir1 (1 mg kg −1 ) + light (iv) and cisplatin (3 mg kg −1 ) (v). Data are presented as mean ± SEM. ** p < 0.01, *** p < 0.001.

Article Snippet: From BioLegend (San Diego, CA), we acquired TruStain FcXTM (anti‐mouse CD16/32, 101 319), APC anti‐mouse CD47 (127 514), APC anti‐mouse CD80 (104 714), PE anti‐mouse CD86 (105 008), FITC anti‐mouse CD3 (100 204), BV421 anti‐mouse CD4 (562 891), PE anti‐mouse CD4 (100 408), FITC anti‐mouse CD206 (141 703), APC anti‐mouse CD8a (162 306), Alexa Fluor® 700 anti‐mouse CD45 (103 128), and FITC anti‐mouse CD11c (117 305), while FITC anti‐SIRPα/SHPS‐1 was purchased from Sino Biological (Beijing, China) and PE anti‐human CD47 (E‐AB‐F1413D) from Elabscience (Wuhan, China).

Techniques: In Vivo, Immunohistochemical staining, Staining, Immunofluorescence

Journal: bioRxiv

Article Title: Bone marrow mesenchymal stromal cells mediate cellular inflammation in HFpEF

doi: 10.1101/2025.03.28.645924

Figure Lengend Snippet: ( A ) Experimental outline of HFpEF in IFNɣ reporter mice (GREAT). ( B ) Flow cytometry gating for IFNɣ expression in CD8+ T cells from visceral adipose tissue. ( C ) Total and IFNɣ-producing CD8+ T cell numbers in visceral adipose tissue (VAT) in GREAT mice with HFpEF compared to GREAT control mice without HFpEF (n=8 mice per group, two-tailed Mann-Whitney test). ( D ) Experimental outline of CD8+ T cell depletion in mice with HFpEF. ( E ) Flow cytometry gating for CD8+ T cells in blood. ( F ) CD8+ T cell numbers in blood and VAT following CD8+ T cell depletion (n=6 per group, unpaired student’s t-test). ( G ) IFNɣ levels in blood in mice with HFpEF with and without CD8+ T cell depletion (n=6 per group, two-tailed Mann-Whitney test). ( H ) CXCL12 levels in the bone marrow plasma of mice with HFpEF with and without CD8+ T cell depletion (n=6 per group, unpaired student’s t-test). ( I ) Gating for blood monocytes in CD8+ depleted mice vs. isotype control mice. ( J ) Monocyte levels in the blood of CD8+ depleted mice vs. isotype controls (n=6 per group, unpaired student’s t-test).

Article Snippet: To deplete CD8+ T cells, mice were intraperitoneally (i.p.) injected with 200 µg of InVivo MAb anti-mouse CD8a (clone 2.43, BioXCell) or InVivo MAb rat IgG2b isotype control, anti-keyhole limpet hemocyanin (clone LTF-2, BioXCell), every 3 days throughout the course of the HFpEF protocol.

Techniques: Flow Cytometry, Expressing, Control, Two Tailed Test, MANN-WHITNEY

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 1. Anti-TGF-b CAR T cells reduced tumor growth and promoted T cell expansion in vivo (A) Anti-TGF-b CAR vector (T28zT2), anti-GPC3 CAR vector (G28zT2), and anti-CD19 CAR vector (1928zT2) based on an anti-TGF-b scFv (US20140127230A1), anti-GPC3 scFv (GC33), or anti-CD19 scFv (FMC63), respectively. All contained expression cassettes encoding a human CD8 leader signal peptide, CD28, CD3z, and TLR2 signaling domain along with eGFP using 2A self-cleaving peptide (2A). The eGFP expression was used to monitor CAR-transduced cells. (B) The percentage of Huh7 cells with 1928zT2, G28zT2, or T28zT2 T cell-induced lysis after 72 h; data are the mean percentage of tumor cell-specific lysis ± SEM values; n = 3 independent experiments; two-way ANOVA with Tukey’s multiple comparisons test; ****p % 0.0001.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb Cell Signaling Technology Cat# 4494 RRID: AB_11178659 OPA1 (D7C1A) Rabbit mAb Cell Signaling Technology Cat# 67589 RRID: AB_2799728 MFF (E5W4M) XP Rabbit mAb Cell Signaling Technology Cat# 84580 RRID: AB_2799819 SMAD4 (D3R4N) XP Rabbit mAb Cell Signaling Technology Cat# 46535 RRID: AB_2736998 SMAD4 Polyclonal antibody Proteintech Cat# 10231-1-AP RRID: AB_2193323 a-Smooth Muscle Actin (D4K9N) XP Rabbit mAb Cell Signaling Technology Cat# 19245 RRID: AB_2734735 Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb Cell Signaling Technology Cat# 9664 RRID: 10828837 CD8a (C8/144B) Mouse mAb Cell Signaling Technology Cat# 70306 RRID: AB_2799781 Anti-CD4 antibody Abcam Cat# ab133616 RRID: AB_2750883 Goat Anti-Rabbit IgG H&L (Alexa Fluor 568) Abcam Cat# ab175471 RRID: AB_2576207 Goat Anti-Mouse IgG H&L (Alexa Fluor 647) Abcam Cat# ab150115 RRID: AB_2687948 Biological samples Patient-derived xenografts (PDX) Jiang. et al.,26 Qin. et al.46 N/A Heathy PBMCs Guangzhou Tianhe Noah Biological Engineering Co., LTD NA Chemicals, peptides, and recombinant proteins TGF beta 1 Protein, Human, Rhesus, Cynomolgus, Canine, Recombinant SinoBiological CAS number: 10804-HNAC TMRM MedChemExpress Cat# HY-D0984 MitoTracker Green Yeasen Cat# 40742ES50 MitoTracker Deep Red Yeasen Cat# 40743ES50 GenCRISPRTM Cas9 v1.2 GenScript Cat# Z03702-1 Seahorse XF Cell Mito Stress Test Kit Agilent Cat# 103010-100 Critical commercial assays Human Granzyme B Precoated ELISA Kit DAKEWE Cat# 1118503 Human IFN-g Precoated ELISA Kit DAKEWE Cat# 1110003 Human IL-2 Precoated ELISA Kit DAKEWE Cat# 1110203 Human TGFb1 Precoated ELISA Kit DAKEWE Cat# 1117102 Human PRF1 Precoated ELISA Kit Bioswamp Cat# HM10300 MACS Pan T cell Isolation Kit Miltenyi Biotec Cat# 130-096-535 MACS GMP T cell TransAct Miltenyi Biotec Cat# 170-076-156 Deposited data Original data of the bulk RNAseq This paper GSA-human: HRA001397 Original western blot images This paper https://doi.org/10.17632/n7htzcxmw4.1 Links: https://data.mendeley.com/drafts/ n7htzcxmw4/1 Experimental models: Cell lines Huh7 Procell Cat# CL-0120 HepG2 Procell Cat# CL-0103 (Continued on next page) Cell Reports Medicine 6, 102020, March 18, 2025 e3

Techniques: In Vivo, Plasmid Preparation, Expressing, Lysis

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 3. CD4+ but not CD8+ T28zT2 T cells are effective for tumor growth inhibition (A) CD4+ and CD8+ T cell compartments were sorted, and CD8+ T28zT2 T cells and CD4+ T28zT2 cells were generated. We also mixed the CD4+ and CD8+ T28zT2 T cells at a ratio of 1:1 to obtain CD3+ T28zT2 T cells. We then infused a total of 5 3 106 of the three types of T28zT2 T cells, CD3+ 1928zT2 T cells, or CD3+ M28zT2 T cells peritumorally into subcutaneous xenografts of NSI mice inoculated with 1 3 106 AsPc-1 cells (day 0). Tumor volumes were monitored on the indicated days

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb Cell Signaling Technology Cat# 4494 RRID: AB_11178659 OPA1 (D7C1A) Rabbit mAb Cell Signaling Technology Cat# 67589 RRID: AB_2799728 MFF (E5W4M) XP Rabbit mAb Cell Signaling Technology Cat# 84580 RRID: AB_2799819 SMAD4 (D3R4N) XP Rabbit mAb Cell Signaling Technology Cat# 46535 RRID: AB_2736998 SMAD4 Polyclonal antibody Proteintech Cat# 10231-1-AP RRID: AB_2193323 a-Smooth Muscle Actin (D4K9N) XP Rabbit mAb Cell Signaling Technology Cat# 19245 RRID: AB_2734735 Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb Cell Signaling Technology Cat# 9664 RRID: 10828837 CD8a (C8/144B) Mouse mAb Cell Signaling Technology Cat# 70306 RRID: AB_2799781 Anti-CD4 antibody Abcam Cat# ab133616 RRID: AB_2750883 Goat Anti-Rabbit IgG H&L (Alexa Fluor 568) Abcam Cat# ab175471 RRID: AB_2576207 Goat Anti-Mouse IgG H&L (Alexa Fluor 647) Abcam Cat# ab150115 RRID: AB_2687948 Biological samples Patient-derived xenografts (PDX) Jiang. et al.,26 Qin. et al.46 N/A Heathy PBMCs Guangzhou Tianhe Noah Biological Engineering Co., LTD NA Chemicals, peptides, and recombinant proteins TGF beta 1 Protein, Human, Rhesus, Cynomolgus, Canine, Recombinant SinoBiological CAS number: 10804-HNAC TMRM MedChemExpress Cat# HY-D0984 MitoTracker Green Yeasen Cat# 40742ES50 MitoTracker Deep Red Yeasen Cat# 40743ES50 GenCRISPRTM Cas9 v1.2 GenScript Cat# Z03702-1 Seahorse XF Cell Mito Stress Test Kit Agilent Cat# 103010-100 Critical commercial assays Human Granzyme B Precoated ELISA Kit DAKEWE Cat# 1118503 Human IFN-g Precoated ELISA Kit DAKEWE Cat# 1110003 Human IL-2 Precoated ELISA Kit DAKEWE Cat# 1110203 Human TGFb1 Precoated ELISA Kit DAKEWE Cat# 1117102 Human PRF1 Precoated ELISA Kit Bioswamp Cat# HM10300 MACS Pan T cell Isolation Kit Miltenyi Biotec Cat# 130-096-535 MACS GMP T cell TransAct Miltenyi Biotec Cat# 170-076-156 Deposited data Original data of the bulk RNAseq This paper GSA-human: HRA001397 Original western blot images This paper https://doi.org/10.17632/n7htzcxmw4.1 Links: https://data.mendeley.com/drafts/ n7htzcxmw4/1 Experimental models: Cell lines Huh7 Procell Cat# CL-0120 HepG2 Procell Cat# CL-0103 (Continued on next page) Cell Reports Medicine 6, 102020, March 18, 2025 e3

Techniques: Inhibition, Generated

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 5. A combination of CD4+ anti-TGF-b CAR T cells and CD8+ anti-MSLN CAR T cells exhibits augmented antitumor effects in NSCLC PDX (A) NSCLC PDX tumors were diced into 30 mm3 pieces, and tissue were inoculated subcutaneously into the right flanks of 8-week-old male NSI mice. 5 3 106

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb Cell Signaling Technology Cat# 4494 RRID: AB_11178659 OPA1 (D7C1A) Rabbit mAb Cell Signaling Technology Cat# 67589 RRID: AB_2799728 MFF (E5W4M) XP Rabbit mAb Cell Signaling Technology Cat# 84580 RRID: AB_2799819 SMAD4 (D3R4N) XP Rabbit mAb Cell Signaling Technology Cat# 46535 RRID: AB_2736998 SMAD4 Polyclonal antibody Proteintech Cat# 10231-1-AP RRID: AB_2193323 a-Smooth Muscle Actin (D4K9N) XP Rabbit mAb Cell Signaling Technology Cat# 19245 RRID: AB_2734735 Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb Cell Signaling Technology Cat# 9664 RRID: 10828837 CD8a (C8/144B) Mouse mAb Cell Signaling Technology Cat# 70306 RRID: AB_2799781 Anti-CD4 antibody Abcam Cat# ab133616 RRID: AB_2750883 Goat Anti-Rabbit IgG H&L (Alexa Fluor 568) Abcam Cat# ab175471 RRID: AB_2576207 Goat Anti-Mouse IgG H&L (Alexa Fluor 647) Abcam Cat# ab150115 RRID: AB_2687948 Biological samples Patient-derived xenografts (PDX) Jiang. et al.,26 Qin. et al.46 N/A Heathy PBMCs Guangzhou Tianhe Noah Biological Engineering Co., LTD NA Chemicals, peptides, and recombinant proteins TGF beta 1 Protein, Human, Rhesus, Cynomolgus, Canine, Recombinant SinoBiological CAS number: 10804-HNAC TMRM MedChemExpress Cat# HY-D0984 MitoTracker Green Yeasen Cat# 40742ES50 MitoTracker Deep Red Yeasen Cat# 40743ES50 GenCRISPRTM Cas9 v1.2 GenScript Cat# Z03702-1 Seahorse XF Cell Mito Stress Test Kit Agilent Cat# 103010-100 Critical commercial assays Human Granzyme B Precoated ELISA Kit DAKEWE Cat# 1118503 Human IFN-g Precoated ELISA Kit DAKEWE Cat# 1110003 Human IL-2 Precoated ELISA Kit DAKEWE Cat# 1110203 Human TGFb1 Precoated ELISA Kit DAKEWE Cat# 1117102 Human PRF1 Precoated ELISA Kit Bioswamp Cat# HM10300 MACS Pan T cell Isolation Kit Miltenyi Biotec Cat# 130-096-535 MACS GMP T cell TransAct Miltenyi Biotec Cat# 170-076-156 Deposited data Original data of the bulk RNAseq This paper GSA-human: HRA001397 Original western blot images This paper https://doi.org/10.17632/n7htzcxmw4.1 Links: https://data.mendeley.com/drafts/ n7htzcxmw4/1 Experimental models: Cell lines Huh7 Procell Cat# CL-0120 HepG2 Procell Cat# CL-0103 (Continued on next page) Cell Reports Medicine 6, 102020, March 18, 2025 e3

Techniques:

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 6. TGF-b1 suppressed OXPHOS activity in CD4+ human T cells (A–D) CD4+ and CD8+ T cells (1 3 106) were activated with CD3/CD28 mAbs for 24 h, followed by PBS or TGF-b1 (10 ng/mL) treatment for 16 h. (A) Mitochondrial morphology of CD4+ and CD8+ T cells upon PBS or TGF-b1 (10 ng/mL) treatment, as determined by spinning disk confocal microscopy. Mitochondria are green (MitoTracker Green), and nuclei are blue (DAPI). Scale bar, 5 mm. (B) Relative lengths of mitochondria, as analyzed by ImageJ software, in CD4+ and CD8+ T cells (2 independent experiments). Each dot represents the mean relative length of the mitochondria in a sample. Data are shown as the mean ± SEM values; paired two-tailed t test; ****p % 0.0001. (C) Immunoblot analysis of cellular protein extracts probed with antibodies against pSMAD2S465/467 (top)/pSMAD3S423/425

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Phospho-DRP1 (Ser616) (D9A1) Rabbit mAb Cell Signaling Technology Cat# 4494 RRID: AB_11178659 OPA1 (D7C1A) Rabbit mAb Cell Signaling Technology Cat# 67589 RRID: AB_2799728 MFF (E5W4M) XP Rabbit mAb Cell Signaling Technology Cat# 84580 RRID: AB_2799819 SMAD4 (D3R4N) XP Rabbit mAb Cell Signaling Technology Cat# 46535 RRID: AB_2736998 SMAD4 Polyclonal antibody Proteintech Cat# 10231-1-AP RRID: AB_2193323 a-Smooth Muscle Actin (D4K9N) XP Rabbit mAb Cell Signaling Technology Cat# 19245 RRID: AB_2734735 Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb Cell Signaling Technology Cat# 9664 RRID: 10828837 CD8a (C8/144B) Mouse mAb Cell Signaling Technology Cat# 70306 RRID: AB_2799781 Anti-CD4 antibody Abcam Cat# ab133616 RRID: AB_2750883 Goat Anti-Rabbit IgG H&L (Alexa Fluor 568) Abcam Cat# ab175471 RRID: AB_2576207 Goat Anti-Mouse IgG H&L (Alexa Fluor 647) Abcam Cat# ab150115 RRID: AB_2687948 Biological samples Patient-derived xenografts (PDX) Jiang. et al.,26 Qin. et al.46 N/A Heathy PBMCs Guangzhou Tianhe Noah Biological Engineering Co., LTD NA Chemicals, peptides, and recombinant proteins TGF beta 1 Protein, Human, Rhesus, Cynomolgus, Canine, Recombinant SinoBiological CAS number: 10804-HNAC TMRM MedChemExpress Cat# HY-D0984 MitoTracker Green Yeasen Cat# 40742ES50 MitoTracker Deep Red Yeasen Cat# 40743ES50 GenCRISPRTM Cas9 v1.2 GenScript Cat# Z03702-1 Seahorse XF Cell Mito Stress Test Kit Agilent Cat# 103010-100 Critical commercial assays Human Granzyme B Precoated ELISA Kit DAKEWE Cat# 1118503 Human IFN-g Precoated ELISA Kit DAKEWE Cat# 1110003 Human IL-2 Precoated ELISA Kit DAKEWE Cat# 1110203 Human TGFb1 Precoated ELISA Kit DAKEWE Cat# 1117102 Human PRF1 Precoated ELISA Kit Bioswamp Cat# HM10300 MACS Pan T cell Isolation Kit Miltenyi Biotec Cat# 130-096-535 MACS GMP T cell TransAct Miltenyi Biotec Cat# 170-076-156 Deposited data Original data of the bulk RNAseq This paper GSA-human: HRA001397 Original western blot images This paper https://doi.org/10.17632/n7htzcxmw4.1 Links: https://data.mendeley.com/drafts/ n7htzcxmw4/1 Experimental models: Cell lines Huh7 Procell Cat# CL-0120 HepG2 Procell Cat# CL-0103 (Continued on next page) Cell Reports Medicine 6, 102020, March 18, 2025 e3

Techniques: Activity Assay, Confocal Microscopy, Software, Two Tailed Test, Western Blot

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 1. Anti-TGF-b CAR T cells reduced tumor growth and promoted T cell expansion in vivo (A) Anti-TGF-b CAR vector (T28zT2), anti-GPC3 CAR vector (G28zT2), and anti-CD19 CAR vector (1928zT2) based on an anti-TGF-b scFv (US20140127230A1), anti-GPC3 scFv (GC33), or anti-CD19 scFv (FMC63), respectively. All contained expression cassettes encoding a human CD8 leader signal peptide, CD28, CD3z, and TLR2 signaling domain along with eGFP using 2A self-cleaving peptide (2A). The eGFP expression was used to monitor CAR-transduced cells. (B) The percentage of Huh7 cells with 1928zT2, G28zT2, or T28zT2 T cell-induced lysis after 72 h; data are the mean percentage of tumor cell-specific lysis ± SEM values; n = 3 independent experiments; two-way ANOVA with Tukey’s multiple comparisons test; ****p % 0.0001.

Article Snippet: Anti-CD8a mouse mAb (clone C8/1448) and a-Smooth Muscle Acitn (a-SMA) (clone D4K9N) rabbit mAb were purchased from Cell Signaling Technology (Boston, USA).

Techniques: In Vivo, Plasmid Preparation, Expressing, Lysis

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 3. CD4+ but not CD8+ T28zT2 T cells are effective for tumor growth inhibition (A) CD4+ and CD8+ T cell compartments were sorted, and CD8+ T28zT2 T cells and CD4+ T28zT2 cells were generated. We also mixed the CD4+ and CD8+ T28zT2 T cells at a ratio of 1:1 to obtain CD3+ T28zT2 T cells. We then infused a total of 5 3 106 of the three types of T28zT2 T cells, CD3+ 1928zT2 T cells, or CD3+ M28zT2 T cells peritumorally into subcutaneous xenografts of NSI mice inoculated with 1 3 106 AsPc-1 cells (day 0). Tumor volumes were monitored on the indicated days

Article Snippet: Anti-CD8a mouse mAb (clone C8/1448) and a-Smooth Muscle Acitn (a-SMA) (clone D4K9N) rabbit mAb were purchased from Cell Signaling Technology (Boston, USA).

Techniques: Inhibition, Generated

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 5. A combination of CD4+ anti-TGF-b CAR T cells and CD8+ anti-MSLN CAR T cells exhibits augmented antitumor effects in NSCLC PDX (A) NSCLC PDX tumors were diced into 30 mm3 pieces, and tissue were inoculated subcutaneously into the right flanks of 8-week-old male NSI mice. 5 3 106

Article Snippet: Anti-CD8a mouse mAb (clone C8/1448) and a-Smooth Muscle Acitn (a-SMA) (clone D4K9N) rabbit mAb were purchased from Cell Signaling Technology (Boston, USA).

Techniques:

Journal: Cell reports. Medicine

Article Title: CD4 + anti-TGF-β CAR T cells and CD8 + conventional CAR T cells exhibit synergistic antitumor effects.

doi: 10.1016/j.xcrm.2025.102020

Figure Lengend Snippet: Figure 6. TGF-b1 suppressed OXPHOS activity in CD4+ human T cells (A–D) CD4+ and CD8+ T cells (1 3 106) were activated with CD3/CD28 mAbs for 24 h, followed by PBS or TGF-b1 (10 ng/mL) treatment for 16 h. (A) Mitochondrial morphology of CD4+ and CD8+ T cells upon PBS or TGF-b1 (10 ng/mL) treatment, as determined by spinning disk confocal microscopy. Mitochondria are green (MitoTracker Green), and nuclei are blue (DAPI). Scale bar, 5 mm. (B) Relative lengths of mitochondria, as analyzed by ImageJ software, in CD4+ and CD8+ T cells (2 independent experiments). Each dot represents the mean relative length of the mitochondria in a sample. Data are shown as the mean ± SEM values; paired two-tailed t test; ****p % 0.0001. (C) Immunoblot analysis of cellular protein extracts probed with antibodies against pSMAD2S465/467 (top)/pSMAD3S423/425

Article Snippet: Anti-CD8a mouse mAb (clone C8/1448) and a-Smooth Muscle Acitn (a-SMA) (clone D4K9N) rabbit mAb were purchased from Cell Signaling Technology (Boston, USA).

Techniques: Activity Assay, Confocal Microscopy, Software, Two Tailed Test, Western Blot

Journal: eBioMedicine

Article Title: Enhancing the efficacy of near-infrared photoimmunotherapy through intratumoural delivery of CD44–targeting antibody–photoabsorber conjugates

doi: 10.1016/j.ebiom.2025.105566

Figure Lengend Snippet: Intratumoural delivery of CD44-IR700 enhanced efficacy of photoimmunotherapy in vivo . (a) Schematic of experimental schedule. Mice bearing subcutaneous Lewis lung carcinoma tumours were treated with either no CD44-IR700 (Ctrl), CD44-IR700 via IV injection (CD44-IR700_IV), or CD44-IR700 via IT injection (CD44-IR700_IT), followed by NIR light irradiation on subsequent days. (b) The ratio of tumour-infiltrating CD8-positive T cells in live cells, as determined by flow cytometry on day 7 post-treatment, from the CD44-IR700_IV and CD44-IR700_IT groups (n = 6 per group). (c) Immunohistochemical staining of tumours 7 days after photoimmunotherapy. CD8-positive T cells were stained. The scale bar represents 50 μm. The numbers of CD8-positive T cells per view field were counted (n = 5 mice for each group). (d) Cytokine expression in tumours, as determined by quantitative PCR analysis on day 3 post-treatment with photoimmunotherapy, from the CD44-IR700_IV and CD44-IR700_IT groups (n = 5 per group). (e) Tumour volume monitoring. Tumour size was measured starting from the day of CD44-IR700 administration (day 0), and the change in size was expressed as a ratio relative to the initial volume on day 0 (n = 12 per group). (f) Tumour volume ratio on day 7. The graph shows the fold increase in tumour volume on day 7 compared with that on day 0 (n = 12 per group). (g) Representative images displaying tumours from each treatment group on day 7 post-administration. (h) Representative images of tumours excised on day 7 post-treatment from each therapeutic group. Data are presented as mean + standard error of the mean. Statistical difference was assessed using Mann–Whitney U tests, with significance denoted by asterisks (∗ p < 0.05 and ∗∗∗∗ p < 0.0001). CD44-IR700, IR700-conjugated anti-CD44 monoclonal antibody; CD, cluster of differentiation; NIR, near-infrared; Ctrl, control; IV, intravenous; IT, intratumoural; Ifnγ, interferon-γ; and Tnf, tumour necrosis factor-alpha.

Article Snippet: For immunohistochemistry, the primary antibody was anti-mouse CD8a antibody (98941S, Cell Signalling Technology, RRID: AB_2756376 ).

Techniques: In Vivo, IV Injection, Injection, Irradiation, Flow Cytometry, Immunohistochemical staining, Staining, Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY, Control