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A subpopulation of RGCs express the stress marker phosphorylated c-JUN <t>(p-cJUN).</t> (A) Whole mount image ( Cx3cr1-Gfp/+ ) of e14.5 retina showing that a subset of RGCs (Brn3a+; red) express nuclear p-cJUN (white; yellow arrows). Boxed regions show enlarged region and label for p-cJUN alone or Brn3a alone (A′) . Scale bar = 10 μm. (B) Whole mount image (e14.5 Cx3cr1-Gfp/+ ) showing microglia (GFP) internalizing an RGC (Brn3a + red) labeled with p-cJUN (white) using HCR IHC. Scale bar = 1 μm. (C) Whole mount images showing RGCs (Brn3a+; red) with nuclear p-cJUN (white) in control mice (WT) and Mertk/CR3 dKO mice. Yellow arrows indicate double positive p-cJUN and Brn3a staining. (D) Quantification of double positive p-cJUN + Brn3a + RGCs per mm 2 in e14.5 control and Mertk/CR3 dKO mice. *** p < 0.01 unpaired t -test [ N = 4 (WT), 5 (Mertk CR3 dKO)]. (E) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in e14.5 and e16.5 Mertk/CR3 dKO mice. * p < 0.05 Welch’s t -test [ N = 5 (e14.5), 4 (e16.5)]. (F) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in P0 control, Mertk KO and Mertk/CR3 dKO mice. ns, not significant. Welch’s one-way ANOVA with Brown–Forsythe variance test [ N = 7 (WT), 7 (Mertk KO), 6 (Mertk CR3 dKO)]. (G) Whole mount retina images to show vehicle treated Axl KO mice with RGCs (Brn3a+; red) expressing nuclear p-cJUN (white) and microglia present (IBA1+; green). Scale bar = 500 μm. Boxed regions show enlarged area (G′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100-10 μm. (H) Whole mount retina images to show Axl KO PLX treated mice with depletion of microglia (IBA1; green) and RGCs (Brn3a+; red) with nuclear p-cJUN (white) expression remaining. Boxed regions show enlarged area (H′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100 to 10 μm.
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A subpopulation of RGCs express the stress marker phosphorylated c-JUN (p-cJUN). (A) Whole mount image ( Cx3cr1-Gfp/+ ) of e14.5 retina showing that a subset of RGCs (Brn3a+; red) express nuclear p-cJUN (white; yellow arrows). Boxed regions show enlarged region and label for p-cJUN alone or Brn3a alone (A′) . Scale bar = 10 μm. (B) Whole mount image (e14.5 Cx3cr1-Gfp/+ ) showing microglia (GFP) internalizing an RGC (Brn3a + red) labeled with p-cJUN (white) using HCR IHC. Scale bar = 1 μm. (C) Whole mount images showing RGCs (Brn3a+; red) with nuclear p-cJUN (white) in control mice (WT) and Mertk/CR3 dKO mice. Yellow arrows indicate double positive p-cJUN and Brn3a staining. (D) Quantification of double positive p-cJUN + Brn3a + RGCs per mm 2 in e14.5 control and Mertk/CR3 dKO mice. *** p < 0.01 unpaired t -test [ N = 4 (WT), 5 (Mertk CR3 dKO)]. (E) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in e14.5 and e16.5 Mertk/CR3 dKO mice. * p < 0.05 Welch’s t -test [ N = 5 (e14.5), 4 (e16.5)]. (F) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in P0 control, Mertk KO and Mertk/CR3 dKO mice. ns, not significant. Welch’s one-way ANOVA with Brown–Forsythe variance test [ N = 7 (WT), 7 (Mertk KO), 6 (Mertk CR3 dKO)]. (G) Whole mount retina images to show vehicle treated Axl KO mice with RGCs (Brn3a+; red) expressing nuclear p-cJUN (white) and microglia present (IBA1+; green). Scale bar = 500 μm. Boxed regions show enlarged area (G′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100-10 μm. (H) Whole mount retina images to show Axl KO PLX treated mice with depletion of microglia (IBA1; green) and RGCs (Brn3a+; red) with nuclear p-cJUN (white) expression remaining. Boxed regions show enlarged area (H′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100 to 10 μm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Microglial mechanisms of viable retinal ganglion cell elimination

doi: 10.3389/fncel.2025.1719791

Figure Lengend Snippet: A subpopulation of RGCs express the stress marker phosphorylated c-JUN (p-cJUN). (A) Whole mount image ( Cx3cr1-Gfp/+ ) of e14.5 retina showing that a subset of RGCs (Brn3a+; red) express nuclear p-cJUN (white; yellow arrows). Boxed regions show enlarged region and label for p-cJUN alone or Brn3a alone (A′) . Scale bar = 10 μm. (B) Whole mount image (e14.5 Cx3cr1-Gfp/+ ) showing microglia (GFP) internalizing an RGC (Brn3a + red) labeled with p-cJUN (white) using HCR IHC. Scale bar = 1 μm. (C) Whole mount images showing RGCs (Brn3a+; red) with nuclear p-cJUN (white) in control mice (WT) and Mertk/CR3 dKO mice. Yellow arrows indicate double positive p-cJUN and Brn3a staining. (D) Quantification of double positive p-cJUN + Brn3a + RGCs per mm 2 in e14.5 control and Mertk/CR3 dKO mice. *** p < 0.01 unpaired t -test [ N = 4 (WT), 5 (Mertk CR3 dKO)]. (E) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in e14.5 and e16.5 Mertk/CR3 dKO mice. * p < 0.05 Welch’s t -test [ N = 5 (e14.5), 4 (e16.5)]. (F) Quantification of p-cJUN + Brn3a + RGCs per mm 2 in P0 control, Mertk KO and Mertk/CR3 dKO mice. ns, not significant. Welch’s one-way ANOVA with Brown–Forsythe variance test [ N = 7 (WT), 7 (Mertk KO), 6 (Mertk CR3 dKO)]. (G) Whole mount retina images to show vehicle treated Axl KO mice with RGCs (Brn3a+; red) expressing nuclear p-cJUN (white) and microglia present (IBA1+; green). Scale bar = 500 μm. Boxed regions show enlarged area (G′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100-10 μm. (H) Whole mount retina images to show Axl KO PLX treated mice with depletion of microglia (IBA1; green) and RGCs (Brn3a+; red) with nuclear p-cJUN (white) expression remaining. Boxed regions show enlarged area (H′) with yellow arrows indicating double positive p-cJUN and Brn3a staining. Scale bars = 100 to 10 μm.

Article Snippet: Antibody , Rabbit anti p-cJUN , Cell Signaling , 1:500 , 9261S.

Techniques: Marker, Labeling, Control, Staining, Expressing