Journal: Journal of Neuroinflammation

Article Title: SIRT2 Inhibition promotes microglia LC3-associated phagocytosis via NRF2/CD36 after the experimental subarachnoid hemorrhage

doi: 10.1186/s12974-025-03623-z

Figure Lengend Snippet: Expression of SIRT2 and its prognostic impact after SAH in rats. A-B, Western blot and quantifications of SIRT2 level at different time courses. C-E, Representative images show in the temporal cortex stained with antibodies to SIRT2 and Iba1 (C), NeuN (D), or GFAP (E) at 24 h post-SAH. F, The mean fluorescence intensity (MFI) of SIRT2 immunoreactivity. G, Percentages of Iba1 + SIRT2 + , NeuN + SIRT2 + and GFAP + SIRT2 + in the temporal cortex of each group. H , Neurological score, I , brain water content ( J ) and Evan’s blue in different (Sham, SAH, Vehicle and AK7) groups at 24 h after SAH. K-L , FJC staining in the cortex with quantification. Scale bar: 50 μm. ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: SIRT2 inhibitor AK7 (MCE, USA) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Expressing, Western Blot, Staining, Fluorescence

Journal: Journal of Neuroinflammation

Article Title: SIRT2 Inhibition promotes microglia LC3-associated phagocytosis via NRF2/CD36 after the experimental subarachnoid hemorrhage

doi: 10.1186/s12974-025-03623-z

Figure Lengend Snippet: SIRT2 inhibition promotes microglia phagocytosis. A-B , Western blot and quantitative analysis of SIRT2 expressions in microglia (Con, OxyHb, DMSO and AK7) groups. C-D ,phagocytic index level and analysis by pHrodo. E-F ,Immunofluorescence staining and quantitative analysis of LC3 and Phrodo in microglia. Scale bar: 10 μm. G-K , Western blot and quantitative analysis of Rubicon, NOX2, Beclin1 and LC3 A/B expressions. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: SIRT2 inhibitor AK7 (MCE, USA) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Inhibition, Western Blot, Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: SIRT2 Inhibition promotes microglia LC3-associated phagocytosis via NRF2/CD36 after the experimental subarachnoid hemorrhage

doi: 10.1186/s12974-025-03623-z

Figure Lengend Snippet: SIRT2 deacetylates NRF2 and promotes its nuclear translocation in microglia. A-C , Heatmap analyses and q-PCR detects different NRF2 target genes in microglia (SIRT2 and SIRT2 KD) groups. D , Molecular docking shown the obvious interaction force between SIRT2 and NRF2. E-G , Co-immunoprecipitation and quantitative analysis of acetylated NRF2 in SIRT2 WT and KD microglia. H , Western blot and quantitative analysis of SIRT2 in microglia (Con, OxyHb, DMSO and AK7) group with Cyto and nucleus. I-K , The co-localization of SIRT2 and nucleus with laser confocal and analysis in Con group and OxyHb group of microglia. Scale bar: 10 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, n.s., no significant

Article Snippet: SIRT2 inhibitor AK7 (MCE, USA) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Translocation Assay, Immunoprecipitation, Western Blot

Journal: Journal of Neuroinflammation

Article Title: SIRT2 Inhibition promotes microglia LC3-associated phagocytosis via NRF2/CD36 after the experimental subarachnoid hemorrhage

doi: 10.1186/s12974-025-03623-z

Figure Lengend Snippet: SIRT2 through NRF2/CD36 regulates microglial phagocytosis. A , JASPAR predict NRF2 bind to the promoter region of CD36. B-C , phagocytic index level and analysis by pHrodo in microglia (Con, OxyHb, DMSO, AK7 and AK7+ML385) groups. D-E , Western blot and quantitative analysis of CD36 expression. F-H , ELISA quantitative analysis of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) level in different groups. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: SIRT2 inhibitor AK7 (MCE, USA) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Neuroinflammation

Article Title: SIRT2 Inhibition promotes microglia LC3-associated phagocytosis via NRF2/CD36 after the experimental subarachnoid hemorrhage

doi: 10.1186/s12974-025-03623-z

Figure Lengend Snippet: SIRT2 via LAP inhibits phagocytosis in microglia. A-F ,Western blot and quantitative analysis of Rubicon, ULK1, CD36, NOX2 and LC3 A/B expressions. G-H , The co-localization of LC3 and pHrodo with laser confocal and analysis indifferent groups of microglia. Scale bar: 10 μm. I-J , Representative TEM images of LAPosomes in microglia. (The black triangle refers to the single membrane structure characteristic of LAPosomes and star refers to the Rubicon particles). Scale bar: 3 μm (upper panels), 1 μm (lower panels). * P < 0.05 vs. Hb, # P < 0.05 vs. Hb+AK7, & P < 0.05 vs. Hb+shRNA, $ P < 0.05 vs. Hb+AK7+shRNA, ☆ P < 0.05 vs. Hb+AK7+ML385, *** P < 0.001, **** P < 0.0001

Article Snippet: SIRT2 inhibitor AK7 (MCE, USA) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Western Blot, Membrane, shRNA

Journal: Journal of Neuroinflammation

Article Title: SIRT2 Inhibition promotes microglia LC3-associated phagocytosis via NRF2/CD36 after the experimental subarachnoid hemorrhage

doi: 10.1186/s12974-025-03623-z

Figure Lengend Snippet: Inhibition of SIRT2 improves long-term neurological function after SAH. A-B , The falling latency of rotarod test (5RPM and 10 RPM) at 1, 2, and 3 weeks after SAH in rats. C-E ,Escape latency and swimming distance of Morris water maze test on days 23 to 27 after SAH. F-G , Quantification of probe quadrant duration and swimming velocities. H-I , Representative images and neuronal quantifications of Nissl staining in CA1 hippocampal region. Scale bar: 200 μm (upper panels), 50 μm (lower panels). * P < 0.05, ** P< 0.01, *** P < 0.001, **** P < 0.0001, n.s., no significant

Article Snippet: SIRT2 inhibitor AK7 (MCE, USA) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Inhibition, Staining

Journal: Journal of Neuroinflammation

Article Title: SIRT2 Inhibition promotes microglia LC3-associated phagocytosis via NRF2/CD36 after the experimental subarachnoid hemorrhage

doi: 10.1186/s12974-025-03623-z

Figure Lengend Snippet: Schematic diagram of our study. SIRT2 modified NRF2 by deacetylation and inhibited its entry into the nucleus, affecting the transcription of downstream CD36, thereby reducing LAP in microglia

Article Snippet: SIRT2 inhibitor AK7 (MCE, USA) was dissolved in dimethyl sulfoxide (DMSO).

Techniques: Modification

Journal: Aging (Albany NY)

Article Title: AK7-deficiency reversal inhibits ccRCC progression and boosts anti-PD1 immunotherapy sensitivity

doi: 10.18632/aging.206006

Figure Lengend Snippet: Expression profile of AK7 in tumor. ( A ) Expression of AK7 mRNA in different cancers and corresponding normal tissues. ( B ) The expression of AK7 mRNA in ccRCC was significantly higher than that in normal kidney tissue. ( C – G ) Differences in SPC25 mRNA expression depending on stage, grade, nodal metastasis status, subtype and race. ( H ) Expression of AK7 in normal renal tissues and ccRCC tissues. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cells were cultured in a 5% CO 2 chamber at 37°C with RFMI-1640 (Gibco, USA) containing 10% FBS (NEWZERUM, Newzerum, Christchurch, New Zealand) and 1% P-S. si-AK7 and si-NC were produced by KeyGEN BioTECH (Jiangsu, China).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: AK7-deficiency reversal inhibits ccRCC progression and boosts anti-PD1 immunotherapy sensitivity

doi: 10.18632/aging.206006

Figure Lengend Snippet: AK7 knockdown promoted the proliferation, invasion and migration ability of human ccRCC cell lines. ( A , B ) Three siRNAs (si1, si2, and si3) were designed to silence AK7 in ccRCC cells (786-O and Caki-1), and validated by qRT-PCR. ( C , D ) The growth curves of 786-O ( C ) and Caki-1 ( D ) cells were plotted after transfection with si1-AK7/si-NC based on CCK-8 assay. ( E , F ) Colony formation assays demonstrated that knockdown of AK7 promoted the proliferation of 786-O and Caki-1 cells. ( G – I ) Transwells experiment demonstrated that knockdown of AK7 expression could effectively promote the migration and invasion ability of ccRCC cells. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cells were cultured in a 5% CO 2 chamber at 37°C with RFMI-1640 (Gibco, USA) containing 10% FBS (NEWZERUM, Newzerum, Christchurch, New Zealand) and 1% P-S. si-AK7 and si-NC were produced by KeyGEN BioTECH (Jiangsu, China).

Techniques: Knockdown, Migration, Quantitative RT-PCR, Transfection, CCK-8 Assay, Expressing

Journal: Aging (Albany NY)

Article Title: AK7-deficiency reversal inhibits ccRCC progression and boosts anti-PD1 immunotherapy sensitivity

doi: 10.18632/aging.206006

Figure Lengend Snippet: Overexpression of AK7 inhibited proliferation, invasion and migration of human ccRCC cell lines. ( A , B ) qRT-PCR verified the efficiency of overexpression of AK7 in 786-O and AKI-1 cell lines. ( C , D ) The growth curves of 786-O ( C ) and Caki-1 ( D ) cells were plotted after overexpression of AK7 based on CCK-8 assay. ( E , F ) Colony formation assays demonstrated that overexpression of AK7 inhibited the proliferation of 786-O and Caki-1 cells. ( G – I ) Transwells experiment demonstrated that overexpression of AK7 could effectively inhibit the migration and invasion ability of ccRCC cells. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cells were cultured in a 5% CO 2 chamber at 37°C with RFMI-1640 (Gibco, USA) containing 10% FBS (NEWZERUM, Newzerum, Christchurch, New Zealand) and 1% P-S. si-AK7 and si-NC were produced by KeyGEN BioTECH (Jiangsu, China).

Techniques: Over Expression, Migration, Quantitative RT-PCR, CCK-8 Assay

Journal: Aging (Albany NY)

Article Title: AK7-deficiency reversal inhibits ccRCC progression and boosts anti-PD1 immunotherapy sensitivity

doi: 10.18632/aging.206006

Figure Lengend Snippet: AK7 can be used as a prognostic indicator and a predictor of immunotherapy effect in ccRCC patients. ( A ) In pancarcinoma, patients with high expression of AK7 have a better prognosis than those with low expression. ( B ) In ccRCC, patients with high expression of AK7 had longer OS than those with low expression. ( C ) In ccRCC at stage 4, patients with high expression of AK7 had longer OS than those with low expression. ( D – F ) In patients treated with anti-PD1 ( D ), anti-PD-L1 ( E ), and anti-CTLA-4 ( F ), high expression of AK7 has a better prognosis. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cells were cultured in a 5% CO 2 chamber at 37°C with RFMI-1640 (Gibco, USA) containing 10% FBS (NEWZERUM, Newzerum, Christchurch, New Zealand) and 1% P-S. si-AK7 and si-NC were produced by KeyGEN BioTECH (Jiangsu, China).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: AK7-deficiency reversal inhibits ccRCC progression and boosts anti-PD1 immunotherapy sensitivity

doi: 10.18632/aging.206006

Figure Lengend Snippet: The expression of AK7 was correlated with immunosuppressive factors. ( A ) Correlation between AK7 and immunoinhibitors in different cancers. ( B ) The expression of AK7 was negatively correlated with the expression of CTLA4, TIGIT, IL10, PD1, IL0RB, LAG3 and other immunosuppressive factors. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cells were cultured in a 5% CO 2 chamber at 37°C with RFMI-1640 (Gibco, USA) containing 10% FBS (NEWZERUM, Newzerum, Christchurch, New Zealand) and 1% P-S. si-AK7 and si-NC were produced by KeyGEN BioTECH (Jiangsu, China).

Techniques: Expressing

Journal: Aging (Albany NY)

Article Title: AK7-deficiency reversal inhibits ccRCC progression and boosts anti-PD1 immunotherapy sensitivity

doi: 10.18632/aging.206006

Figure Lengend Snippet: Overexpression of AK7 inhibits RCC growth and enhances anti-PD1 efficacy. ( A ) Schematic diagram of establishment of mouse subcutaneous tumor model in each group. ( B ) Three lentiviruses were designed to overexpress AK7 expression in RENCA cell lines, and their overexpression efficiency was verified by qRT-PCR. ( C ) Images of subcutaneous tumors in each group. ( D , E ) Analysis of subcutaneous tumors in the respective groups. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cells were cultured in a 5% CO 2 chamber at 37°C with RFMI-1640 (Gibco, USA) containing 10% FBS (NEWZERUM, Newzerum, Christchurch, New Zealand) and 1% P-S. si-AK7 and si-NC were produced by KeyGEN BioTECH (Jiangsu, China).

Techniques: Over Expression, Expressing, Quantitative RT-PCR