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(A) Representative confocal immunofluorescence images showing ApoE and PLIN2 staining (green). Scale bar = 30 µm. Lamp2a -/- mice showed increased staining of both proteins compared with WT mice, which was markedly reduced by GLA-1-1 treatment. (B) Bar graph showing quantification of ApoE fluorescence intensity (mean gray value) (n = 7 mice). (C) Bar graph showing quantification of PLIN2 immunofluorescence signal (n = 5 mice). (D) Representative confocal immunofluorescence images showing staining for <t>vitronectin,</t> MMP2, and clusterin. Scale bar = 30 µm. Compared with WT mice, Lamp2a -/- mice showed markedly increased immunoreactivity for these proteins, which was significantly reduced by GLA-1-1 treatment. (E) Bar graph showing quantification of MMP2 fluorescence intensity (mean gray value) (n=5 mice). (F) Bar graph showing quantification of vitronectin fluorescence intensity (mean gray value) (n=7 mice). (G) Bar graph showing quantification of clusterin fluorescence intensity (mean gray value) (n=7 mice). (H) Representative immunoblots showing protein levels of ApoE, clusterin, vitronectin, and PLIN2 in RPE/choroid lysates. Bar graphs showing immunoblot quantification of protein levels of vitronectin (I), clusterin (J), ApoE (K), and PLIN2 (L) in RPE/choroid lysates (n = 3 mice per group). Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Values are expressed as mean ± SD.
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(A) Representative confocal immunofluorescence images showing ApoE and PLIN2 staining (green). Scale bar = 30 µm. Lamp2a -/- mice showed increased staining of both proteins compared with WT mice, which was markedly reduced by GLA-1-1 treatment. (B) Bar graph showing quantification of ApoE fluorescence intensity (mean gray value) (n = 7 mice). (C) Bar graph showing quantification of PLIN2 immunofluorescence signal (n = 5 mice). (D) Representative confocal immunofluorescence images showing staining for <t>vitronectin,</t> MMP2, and clusterin. Scale bar = 30 µm. Compared with WT mice, Lamp2a -/- mice showed markedly increased immunoreactivity for these proteins, which was significantly reduced by GLA-1-1 treatment. (E) Bar graph showing quantification of MMP2 fluorescence intensity (mean gray value) (n=5 mice). (F) Bar graph showing quantification of vitronectin fluorescence intensity (mean gray value) (n=7 mice). (G) Bar graph showing quantification of clusterin fluorescence intensity (mean gray value) (n=7 mice). (H) Representative immunoblots showing protein levels of ApoE, clusterin, vitronectin, and PLIN2 in RPE/choroid lysates. Bar graphs showing immunoblot quantification of protein levels of vitronectin (I), clusterin (J), ApoE (K), and PLIN2 (L) in RPE/choroid lysates (n = 3 mice per group). Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Values are expressed as mean ± SD.
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(A) Representative confocal immunofluorescence images showing ApoE and PLIN2 staining (green). Scale bar = 30 µm. Lamp2a -/- mice showed increased staining of both proteins compared with WT mice, which was markedly reduced by GLA-1-1 treatment. (B) Bar graph showing quantification of ApoE fluorescence intensity (mean gray value) (n = 7 mice). (C) Bar graph showing quantification of PLIN2 immunofluorescence signal (n = 5 mice). (D) Representative confocal immunofluorescence images showing staining for vitronectin, MMP2, and clusterin. Scale bar = 30 µm. Compared with WT mice, Lamp2a -/- mice showed markedly increased immunoreactivity for these proteins, which was significantly reduced by GLA-1-1 treatment. (E) Bar graph showing quantification of MMP2 fluorescence intensity (mean gray value) (n=5 mice). (F) Bar graph showing quantification of vitronectin fluorescence intensity (mean gray value) (n=7 mice). (G) Bar graph showing quantification of clusterin fluorescence intensity (mean gray value) (n=7 mice). (H) Representative immunoblots showing protein levels of ApoE, clusterin, vitronectin, and PLIN2 in RPE/choroid lysates. Bar graphs showing immunoblot quantification of protein levels of vitronectin (I), clusterin (J), ApoE (K), and PLIN2 (L) in RPE/choroid lysates (n = 3 mice per group). Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Values are expressed as mean ± SD.

Journal: bioRxiv

Article Title: Loss of Lamp2a-dependent chaperone-mediated autophagy drives dry AMD-like retinal pathology in mice and is rescued by BK channel activation

doi: 10.64898/2026.03.19.712761

Figure Lengend Snippet: (A) Representative confocal immunofluorescence images showing ApoE and PLIN2 staining (green). Scale bar = 30 µm. Lamp2a -/- mice showed increased staining of both proteins compared with WT mice, which was markedly reduced by GLA-1-1 treatment. (B) Bar graph showing quantification of ApoE fluorescence intensity (mean gray value) (n = 7 mice). (C) Bar graph showing quantification of PLIN2 immunofluorescence signal (n = 5 mice). (D) Representative confocal immunofluorescence images showing staining for vitronectin, MMP2, and clusterin. Scale bar = 30 µm. Compared with WT mice, Lamp2a -/- mice showed markedly increased immunoreactivity for these proteins, which was significantly reduced by GLA-1-1 treatment. (E) Bar graph showing quantification of MMP2 fluorescence intensity (mean gray value) (n=5 mice). (F) Bar graph showing quantification of vitronectin fluorescence intensity (mean gray value) (n=7 mice). (G) Bar graph showing quantification of clusterin fluorescence intensity (mean gray value) (n=7 mice). (H) Representative immunoblots showing protein levels of ApoE, clusterin, vitronectin, and PLIN2 in RPE/choroid lysates. Bar graphs showing immunoblot quantification of protein levels of vitronectin (I), clusterin (J), ApoE (K), and PLIN2 (L) in RPE/choroid lysates (n = 3 mice per group). Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Values are expressed as mean ± SD.

Article Snippet: Antibodies against LC3B (Sigma, L8918-200UL), actin (Proteintech, 66009-1-Ig), PLIN2 (Proteintech, 15294-1-AP), fibronectin (Invitrogen, PA5-29578), clusterin (R&D systems, AF2747), vitronectin (R&D Systems, MAB38751), and p62 (Abnova, H00008878-M01; CST, 5114), Lamp2a (Abcam, ab125068), Lamp2b (Abcam, ab13524), Iba1 (Fujifilm Wako, 019-19741), Laminin (Sigma, L9393) and ApoE (Abcam, ab183596) were used in this study.

Techniques: Immunofluorescence, Staining, Fluorescence, Western Blot