Journal: Molecular Biomedicine

Article Title: TBC1 domain family member 23 is essential for STING-mediated anti-melanoma effect

doi: 10.1186/s43556-026-00418-3

Figure Lengend Snippet: High TBC1D23 expression is correlated with greater immune cell infiltration in melanoma. a Expression level of TBC1D23 in melanoma (n = 558) and adjacent normal (n = 461) tissues from TCGA database was analyzed using the GEPIA 2 on 12 October, 2024 ( http://gepia2.cancer-pku.cn/#analysis ). Bars indicate the median expression level in each group. Statistical analysis was performed using t test. b Prognostic value of TBC1D23 in melanoma was analyzed using the Kaplan–Meier Plotter database ( http://kmplot.com/analysis/ ), which integrates multiple GEO datasets. Overall survival, event free survival and post progression survival were analyzed in all patients with melanoma. Log-rank tests were performed. HR below 1 implies greater survival probabilities for TBC1D23 high group compared with TBC1D23 low group. Detailed information on tumor samples can be found in the individual GEO dataset in the NCBI GEO website. c The estimated enrichment of 7 immune cells in the TBC1D23 high and TBC1D23 low groups was calculated by CIBERSORT. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. d Kaplan–Meier plots for CD8 + T cells infiltrates and TBC1D23 mRNA expression to visualize survival differences in melanoma. e Representative images of mIHC staining of the expression pattern of TBC1D23 and CD8a in human melanoma tissue microarrays. Scale bar 50 μm. f Correlation analysis between the number of TBC1D23 positive cells and the number of CD8 + T cells in human melanoma tissue microarrays. Human melanoma tissue microarrays were generated by SHANGHAI OUTDO BIOTECH CO.LTD. Data were analyzed by Pearson’s correlation

Article Snippet: CD8+ T cells were isolated from spleens and lymph nodes of wild-type C57BL/6 mice using CD8+ T cell isolation beads (MCE #HY-K0310), then activated by stimulation for 72 h on plates coated with 1 μg/mL anti-CD3 and 1 μg/mL anti-CD28, supplemented with 20 ng/mL mIL-2.

Techniques: Expressing, Staining, Generated

Journal: Molecular Biomedicine

Article Title: TBC1 domain family member 23 is essential for STING-mediated anti-melanoma effect

doi: 10.1186/s43556-026-00418-3

Figure Lengend Snippet: TBC1D23 is critical for STING-dependent infiltration of immune cells. a , b Representative FACS data of frequency ( a ) and quantification ( b ) of tumor infiltrating CD45 + T cells or CD8 + T cells (n = 5 per group) of the mice as in Fig. a. c B16F10 tumors were isolated as in Fig. a, then stained with anti-CD45-cy3, anti-F4/80-cy5, anti-CD8-spAqua or anti-GZMB-FITC, and counterstained with DAPI. F4/80 is marker of macrophages. Scale bar, 50 μm. d Quantitative analysis of the numbers of CD45 + cells, macrophages, CD8 + T cells and GZMB + CD8. + T cells within same area of tumors as in c. Data are representative of three independent experiments. means ± SEM. Data were analyzed by one-way ANOVA ( c and d )

Article Snippet: CD8+ T cells were isolated from spleens and lymph nodes of wild-type C57BL/6 mice using CD8+ T cell isolation beads (MCE #HY-K0310), then activated by stimulation for 72 h on plates coated with 1 μg/mL anti-CD3 and 1 μg/mL anti-CD28, supplemented with 20 ng/mL mIL-2.

Techniques: Isolation, Staining, Marker

Journal: Molecular Biomedicine

Article Title: TBC1 domain family member 23 is essential for STING-mediated anti-melanoma effect

doi: 10.1186/s43556-026-00418-3

Figure Lengend Snippet: Deletion of TBC1D23 in macrophages impairs STING-mediated antigen presentation. a , b Mean fluorescent intensity (MFI) of MHC-II (A) and CD86 (B) in BMDMs stimulated with or without DMXAA. WT and Tbc1d23 KO BMDMs were stimulated with or without DMXAA for 24 h, then subjected to flow cytometry analysis (n = 3 biologically independent samples). Left and right panels show representative histograms and quantitative data, respectively. c , d Flow cytometric analysis of expression of H2-k b and H2-k b -SIINFEKL on the surface of B16F10-OVA. WT and Tbc1d23 KO BMDMs were treated with or without DMXAA, supernatants were collected and applied to B16F10-OVA cells pre-treated with either isotype control or anti-Ifnar1. Expression of H2-k b and H2-k b -SIINFEKL on the surface of B16F10-OVA was analyzed by flow cytometry. Left and right panels show representative histograms and quantitative data, respectively. e–g Flow cytometric analysis of antigen-specific cytotoxicity in CD8⁺ T cells. WT and Tbc1d23 KO BMDMs were treated with or without DMXAA, supernatants were collected and then applied to B16F10-OVA cells. CD8 + T cells were isolated from the spleen and lymph node of OT-I mice and co-cultured with B16F10-OVA cells pretreated with supernatants for 48 h. Percentages of GZMB + CD8 + T cells in CD8 + T cells ( e ), and CD107a + CD8 + T cells in CD8 + T cells ( f ), Zombie NIR. + cells in tumor cells ( g ) were analyzed by flow cytometry. Data are representative of three independent experiments. Data were analyzed by one-way ANOVA ( a–d , g ) or by unpaired t test ( e–f )

Article Snippet: CD8+ T cells were isolated from spleens and lymph nodes of wild-type C57BL/6 mice using CD8+ T cell isolation beads (MCE #HY-K0310), then activated by stimulation for 72 h on plates coated with 1 μg/mL anti-CD3 and 1 μg/mL anti-CD28, supplemented with 20 ng/mL mIL-2.

Techniques: Immunopeptidomics, Flow Cytometry, Expressing, Control, Isolation, Cell Culture