anti mouse sirpα pe antibody  (Miltenyi Biotec)

Journal: Science translational medicine

Article Title: Hematopoietic loss of Y chromosome activates immune checkpoints and contributes to impaired senescent cell clearance and renal disease

doi: 10.1126/scitranslmed.adv4071

Figure Lengend Snippet: ( A ) Flow cytometric detection of immune inhibitory receptors (left, PD-1; right, SIRPα) on the indicated immune cell types in kidneys of aged LOY and CT mice at 15 months after BMT (CT: n = 3; LOY: n = 3). Statistical significance was determined by a two-way ANOVA with Tukey’s post hoc test. ( B ) Flow cytometric analyses for PD-1 and SIRPα immune inhibitory receptor expression on the indicated immune cell types in injured kidneys of LOY and CT mice at 4 weeks after the administration of AA (CT: n = 6; LOY: n = 5). Statistical significance was determined by unpaired Student’s t tests. ( C ) Schematic of experiment assessing kidney injury in the hematopoietic LOY model. A series of peritoneal injections every 3 days with combined PD-1 and SIRPα ICI antibodies in LOY and CT mice was started 1 week after the administration of AA, and renal phenotypes were evaluated 4 weeks after the administration of AA. ( D ) Serum BUN amounts (CT control: n = 6; LOY control: n = 5; CT ICIs: n = 6; LOY ICIs: n = 6). Statistical significance was determined by a one-way ANOVA with Tukey’s post hoc test. ( E ) qRT-PCR analysis of kidney injury marker genes in kidneys (CT control: n = 6; LOY control: n = 5; CT ICIs: n = 6; LOY ICIs: n = 6). Statistical significance was determined by unpaired Student’s t tests. ( F ) qRT-PCR analysis of senescence-associated genes in kidneys (CT control: n = 6; LOY control: n = 5; CT ICIs: n = 6; LOY ICIs: n = 6). Statistical significance was determined by a one-way ANOVA with Tukey’s post hoc test. ( G ) Representative images and quantitative analysis of senescent area in kidneys, as stained with SA-β-gal (CT control: n = 6; LOY control: n = 5; CT ICIs: n = 6; LOY ICIs: n = 6). Scale bars, 50 μm. Statistical significance was determined by unpaired Student’s t tests. ( H ) qRT-PCR analysis of the fibrosis-related gene Acta2 in kidneys (CT control: n = 6; LOY control: n = 5; CT ICIs: n = 6; LOY ICIs: n = 6). Statistical significance was determined by a one-way ANOVA with Tukey’s post hoc test. ( I ) Representative images and quantitative analysis of fibrotic area in kidneys, as stained with picrosirius red (CT control: n = 6; LOY control: n = 5; CT ICIs: n = 6; LOY ICIs: n = 6). Scale bars, 50 μm. Statistical significance was determined by unpaired Student’s t tests. Dots in all panels represent individual samples. Data are shown as the mean ± SEM (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and NS ≥ 0.05).

Article Snippet: To evaluate the effect of treatment with ICIs on LOY-mediated renal damage in the murine model of AA-induced fibrosis and senescence, an anti–PD-1 antibody (Bio X Cell, no. BE0146) and an anti-SIRPα antibody (Bio X Cell, no. BE0322) were administered to hematopoietic mosaic LOY and CT mice 1 week after the administration of AA.

Techniques: Expressing, Control, Quantitative RT-PCR, Marker, Staining