Journal: NPJ Vaccines

Article Title: A KIF20A-based thermosensitive hydrogel vaccine effectively potentiates immune checkpoint blockade therapy for hepatocellular carcinoma

doi: 10.1038/s41541-024-01060-2

Figure Lengend Snippet: a CD80 and CD86 levels of BMDCs in flow cytometry (The gating strategy was shown in the Supplementary Fig. ). b Quantification of the proportion of CD80 + CD86 + BMDCs. c MHC-I level of BMDCs indicated by flow cytometry (The gating strategy was shown in the Supplementary Fig. ). d Quantification of the proportion of MHC-I + BMDCs. e MHC-II level of BMDCs in flow cytometry (The gating strategy was shown in the Supplementary Fig. ). f Quantification of MHC-II + BMDCs. g CFDA-SE level in CD3 + T cells (The gating strategy was shown in the Supplementary Fig. ). h Quantification of T cell proliferation rate. i CLSM images of infiltrated CD8a + T cells into Hepa1-6 cell spheroids. j Quantification of T cell infiltration.

Article Snippet: The cells were then stained with APC-conjugated anti-mouse CD3 antibody (Elabscience, China), and the CFDA-SE fluorescence intensity of CD3 + cells was analyzed by flow cytometry to assess T cell proliferation rate.

Techniques: Flow Cytometry

Journal: NPJ Vaccines

Article Title: A KIF20A-based thermosensitive hydrogel vaccine effectively potentiates immune checkpoint blockade therapy for hepatocellular carcinoma

doi: 10.1038/s41541-024-01060-2

Figure Lengend Snippet: a Scheme illustration of in vivo treatment and monitoring (Created by the authors with Photoshop v22.5.6). b Bioluminescence images of tumor-bearing mice in different groups. c Relative signal intensity of tumors in different groups. d Body weight of the mice in different groups. e Flow cytometry analyzing CD80 and CD86 in CD11c + dendritic cells in mouse inguinal lymph nodes (Gating strategy was shown in the Supplementary Fig. ). f Quantification of the percentage of CD80 + CD86 + cells in CD11c + dendritic cells. g Flow cytometry analyzing CD8a and CD4 of T cells (CD45 + CD3 + ) in tumor tissues (Gating strategy was shown in the Supplementary Fig. ). h Quantification of the percentage of CD8a + cells in T cells.

Article Snippet: The cells were then stained with APC-conjugated anti-mouse CD3 antibody (Elabscience, China), and the CFDA-SE fluorescence intensity of CD3 + cells was analyzed by flow cytometry to assess T cell proliferation rate.

Techniques: In Vivo, Flow Cytometry

Journal: Cell reports

Article Title: Epithelial HMGB1 Delays Skin Wound Healing and Drives Tumor Initiation by Priming Neutrophils for NET Formation.

doi: 10.1016/j.celrep.2019.10.104

Figure Lengend Snippet: Figure 5. Keratinocyte-Specific HMGB1 Mediates Regenerative and Tumorigenic Responses through TNF and RIPK1 Kinase Activity (A) Paraffin sections of wounded and tumor-bearing InvEE HMGB1fl/fland DKer HMGB1 skin were labeled with an anti-HMGB1 (red) antibody and counterstained with DAPI (blue). Scale bars: 200 mm. dpw, days post-wounding. Arrowheads indicate the wound edge. (B) Serum levels of HMGB1 in unwounded, wounded (d2pw, day 2 post-wounding), inflamed (InvEE), and InvEE mice bearing a wound-induced papilloma, as assessed by ELISA (data represent mean ± SEM; n R 4 per condition; *p < 0.05; **p = 0.0012; Mann-Whitney test). (C) Western blotting for HMGB1 and H3citr in skin lysates of unwounded (unw), and wounded (day 2, 6, and 8 post-wounding [pw]) InvEE HMGB1fl/fl(wild-type [WT]) and DKer HMGB1 (knockout [KO]) mice. a-actin blot is shown as loading control. (D) Concentration of TNF, IL-6, and IL-8 in serum of unwounded and wounded InvEE HMGB1fl/fl(WT) and DKer HMGB1 (KO) mice (data represent mean ± SEM; n R 4 per condition; *p = 0.0317; Mann-Whitney test). (E) Staining for TNF in wounded skin of InvEE HMGB1fl/fland DKer HMGB1 mice at day 2 post-wounding. Scale bars: 50 mm. (F) Wound-induced tumor incidence in InvEE HMGB1fl/flmice intradermally injected with control IgG (n = 10) or TNF antagonistic antibody (n = 10) at time of wounding and 12, 24, and 36 h post-wounding (**p = 0.002; Wilcoxon matched-pairs signed rank test). (G) Western blotting for H3citr of day 2 post-wounding skin lysates of DKer HMGB1 and HMGB1fl/flmice injected with a-TNF or a-IgG control antibodies. a-actin blot is shown as loading control. (H) Wound-induced skin tumor incidence in InvEE RIPK1WT/WT mice (n = 10) and InvEE RIPK1D138N/D138N mice (n = 10) (**p = 0.0039; Wilcoxon matched-pairs signed rank test). (I) Western blotting for H3citr of day 2 post-wounding skin lysates of InvEE RIPK1D138N/D138N and InvEE RIPK1WT/WT mice. a-actin blot is shown as loading control.

Article Snippet: In conditions where mice were treated, animals were intradermally injected with Box A (250 mg), ethylpyruvate (40 mg/kg), DNase-1 (1000 units), TNF blocking antibody (MP6-XT22 Mab; 100 mg/injection/ mouse), RAGE antagonistic peptide (R&D Systems: 100 mg/injection), or TLR-4 blocking antibody (Novimmune, clone 5E3; 100 mg/injection) in 200 mL PBS, respectively, at time of wounding and at 12, 24 and 36 hours after wounding.

Techniques: Activity Assay, Labeling, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Western Blot, Knock-Out, Control, Concentration Assay, Staining, Injection