Journal: Nature Communications

Article Title: Myeloid EGFR deficiency accelerates recovery from AKI via macrophage efferocytosis and neutrophil apoptosis

doi: 10.1038/s41467-025-59393-y

Figure Lengend Snippet: NeutEGFR −/− and WT mice underwent ischemic injury and were sacrificed at the indicated time points. A Three days after ischemic injury, expression of both p-EGFR and Mcl-1 in kidney neutrophils was evident in WT mice but was minimal in NeutEGFR −/− mice. B – D After ischemic injury, the number of Mcl-1 expressing neutrophils was comparable between WT and NeutEGFR −/− mice at 2 h and 6 h but was markedly lower at 16 and 72 h in NeutEGFR −/− mice ( n = 5–8). E In isolated WT bone marrow neutrophils EGF administration for 16 h induced immunoreactive Mcl-1 protein, which was markedly attenuated in EGFR −/− bone marrow neutrophils isolated from myeloid EGFR −/− mice. Scale bar = 50 μm for all. Data are means ± SD, ** P < 0.01, *** P < 0.001, analyzed using 2-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: The Mcl-1 inhibitor S63845 (MedChemExpress) was formulated and protected from light in 2% Vitamin E/TPGS (Sigma) in NaCl 0.9% (w/v) and was delivered to wild-type C57BL/6 mice by retro-orbital injection for 2 days prior to, and on the day of surgery for acute experiments.

Techniques: Expressing, Isolation

Journal: Nature Communications

Article Title: Myeloid EGFR deficiency accelerates recovery from AKI via macrophage efferocytosis and neutrophil apoptosis

doi: 10.1038/s41467-025-59393-y

Figure Lengend Snippet: WT mice were pretreated with the Mcl-1 inhibitor, S63845 and sacrificed 16 h after the ischemic insult. A – D S63845 treatment led to decreases in numbers of kidney neutrophils but increases in neutrophil apoptosis evaluated with flow cytometry analysis of cleaved caspase 3 ( n = 4) ( A ) and cleaved caspase-9 and S100A9 co-staining (4 and 5) ( B ), in association with less injury, indicated by lower BUN ( n = 6 and 7) ( C ) and a lower tubular injury score ( n = 7 and 8) ( D ). E Mcl-1 inhibition with S63845 led to increased ex vivo bone marrow neutrophil apoptosis determined by cleaved caspase-3 flow cytometric analysis ( n = 4). Of note, cleaved caspase-3 neutrophil counts vs. total neutrophil counts are presented in parenthesis. Scale bar: 50 μm for ( B ) and 100 μm for ( D ). Data are means ± SEM, * P < 0.05, ** P < 0.01, analyzed using 2 tailed Student’s t test for ( A – D ); 2-way ANOVA followed by Tukey’s post hoc test for ( E ).

Article Snippet: The Mcl-1 inhibitor S63845 (MedChemExpress) was formulated and protected from light in 2% Vitamin E/TPGS (Sigma) in NaCl 0.9% (w/v) and was delivered to wild-type C57BL/6 mice by retro-orbital injection for 2 days prior to, and on the day of surgery for acute experiments.

Techniques: Flow Cytometry, Staining, Inhibition, Ex Vivo

Journal: Nature Communications

Article Title: Myeloid EGFR deficiency accelerates recovery from AKI via macrophage efferocytosis and neutrophil apoptosis

doi: 10.1038/s41467-025-59393-y

Figure Lengend Snippet: A Male C57BL/6 (8 weeks old) mice underwent ischemic injury (renal pedicle clamping, 35 min, both kidneys, Bi-IRI)), and the Mcl-1 inhibitor ( S63845 ) was given 6 h later and daily for the next 5 days via intravenous injection at a dose of 5 mg/kg. The mice were sacrificed 3 weeks later. Mcl-1 inhibition accelerated functional recovery, indicated by: B rapid BUN decline ( n = 9 and 10), C preserved GFR ( n = 8) and attenuation of the development of kidney fibrosis, indicated by: D lower mRNA levels ( n = 6 and 9) and E protein levels ( n = 7) of profibrotic and fibrotic components as well as F quantitative Picrosirius red staining ( n = 6) ( F ). Scale bar=100 μm. Data are means ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, analyzed using 2 tailed Student’s t test for all.

Article Snippet: The Mcl-1 inhibitor S63845 (MedChemExpress) was formulated and protected from light in 2% Vitamin E/TPGS (Sigma) in NaCl 0.9% (w/v) and was delivered to wild-type C57BL/6 mice by retro-orbital injection for 2 days prior to, and on the day of surgery for acute experiments.

Techniques: Injection, Inhibition, Functional Assay, Staining