Journal: Cell reports

Article Title: SIRT2 suppresses aging-associated cGAS activation and protects aged mice from severe COVID-19

doi: 10.1016/j.celrep.2025.115562

Figure Lengend Snippet: Young (5 months old) or old (24 months old) mice were inoculated intranasally with control or ORF3a-expressing AAV6. Mice were analyzed 3 weeks later. n =2 independent experiments. (A and B) Quantitative real-time PCR analyses of the mRNA levels of the indicated genes in BALF cells of young and old mice. n = 8. (C) Quantitative real-time PCR analyses of the mRNA levels of IFNβ in the lungs of young mice. n = 6. (D and E) H&E staining (D) and quantification (E) of lung sections of young mice. Scale bar: 100 μm n = 6 mice/group, 10–15 images examined from 3 slides/mouse. (F) Flow cytometry analysis of myeloid cells (Gr1 + CD11b + ) in BALF of young mice. n = 6. (G and H) Immunostaining for Ly6G + cells (G) and quantification (H) of lung sections. Scale bar: 100 μm n = 8 mice/group, 6 images examined from 3 slides/mouse. (I and J) Immunostaining for CD68 + cells (I) and quantification (J) of lung sections. Scale bar: 100 μm n = 8 mice/group, 4 images examined from 3 slides/mouse. (K and L) Sirius red staining (K) and quantification (L) of lung sections. Scale bar: 200 μm n = 6 mice/group, 10 images examined from 3 slides/mouse. (M and N) Immunostaining for phosphorylated STING (M) and quantification (N) of lung sections of young and old mice. Scale bar: 30 μm n = 8 mice/group, 5 images examined from 3 slides/mouse. Data are mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. See also – .

Article Snippet: CD68 clone FA-11 Rat monoclonal antibody , Bio-Rad , Cat# MCA1957GA; RRID:AB_324217.

Techniques: Control, Expressing, Real-time Polymerase Chain Reaction, Staining, Flow Cytometry, Immunostaining

Journal: Investigative Ophthalmology & Visual Science

Article Title: Nicotinamide Riboside Mitigates Retinal Degeneration by Suppressing Damaged DNA-Stimulated Microglial Activation and STING-Mediated Pyroptosis

doi: 10.1167/iovs.66.4.14

Figure Lengend Snippet: Antibodies for Immunofluorescence

Article Snippet: CD68 (FA-11) , Rat , 1:200 , Thermo , 14-0681-82.

Techniques:

Journal: Investigative Ophthalmology & Visual Science

Article Title: Nicotinamide Riboside Mitigates Retinal Degeneration by Suppressing Damaged DNA-Stimulated Microglial Activation and STING-Mediated Pyroptosis

doi: 10.1167/iovs.66.4.14

Figure Lengend Snippet: NR treatment inhibited apoptosis and microglial activation in the LIRD mouse model. (A) Representative morphologic images of each group, with TUNEL (red) labeling apoptotic cells and DAPI ( blue ) staining cell nuclei. Scale bar : 50 µm. (B) Quantification of TUNEL-positive nuclei in the ONL, counted from the entire retina ( n = 5–6/group). Mice treated with NR exhibited significantly fewer TUNEL-positive cells compared to the PBS-treated group. (C) Representative images of Iba1-positive and CD68-positive (phagocytic microglia marker) cells in naive, PBS+LIRD3d, and NR+LIRD3d groups. Scale bar : 50 µm. (D) Quantification of Iba1-positive cells and (E) CD68-positive cells in both ONL and whole retina for each group at three days after light exposure ( n = 4/group). NR treatment significantly suppressed microglia activation, compared with the PBS-treated group ( n = 4/group). * P < 0.05, ** P < 0.01, **** P < 0.0001 by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: CD68 (FA-11) , Rat , 1:200 , Thermo , 14-0681-82.

Techniques: Activation Assay, TUNEL Assay, Labeling, Staining, Marker

Journal: Investigative Ophthalmology & Visual Science

Article Title: Nicotinamide Riboside Mitigates Retinal Degeneration by Suppressing Damaged DNA-Stimulated Microglial Activation and STING-Mediated Pyroptosis

doi: 10.1167/iovs.66.4.14

Figure Lengend Snippet: STING deficiency attenuated light-induced microglial activation and protected against retinal degeneration. (A) Representative images of entire retinal section co-stained with Iba1 ( green ), STING ( red ), and CD68 ( yellow ) from LIRD mice with or without H151 treatment. Scale bar : 500 µm. Magnified single-plane images were shown on the right. Scale bar : 100 µm. (B) Quantification of Iba1+ cells, STING+ Iba1+ cells, and CD68+ Iba1+ cells of each group in ONL at three days after light exposure ( n = 5/group). Phagocytic microglia (CD68+ Iba1+ cells) migrated to the ONL after light exposure, whereas STING deficiency reduced light-induced migration of phagocytic microglia. Arrows point to the colocalization of markers. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, with unpaired t -test. (C) Colocalization of pyroptosis-related proteins (red) and Iba1+ microglia (green) in the retinal sections from each group. Scale bar : 30 µm. (D) Quantification of NLRP3+, Caspase-1+, GSDMD+, and IL-1β+ in Iba1+ cells in ONL at three days after light exposure ( n = 5/group). (E) Representative ERG waveforms of a single eye from each group. (F) Scotopic ERG a-wave ( left panel ) and b-wave ( right panel ) mean amplitudes from LIRD mice treated with H151 or combination of H151 and NR ( n = 4–5/group). Mice treated with H151 exhibited significantly higher a- and b-wave mean amplitudes ( blue line ) compared to the LIRD group ( orange line ). NR supplement ( black line ) did not add more protection to H151-treated retinas. Compared with the H151+LIRD group, * P < 0.05; Compared with the H151+NR+LIRD group, # P < 0.05, ## P < 0.01, #### P < 0.0001, by two-way ANOVA with Tukey's multiple comparisons test. Error bars : SEM.

Article Snippet: CD68 (FA-11) , Rat , 1:200 , Thermo , 14-0681-82.

Techniques: Activation Assay, Staining, Migration