In vivo ER-phagy is sensitive to physio-pathological perturbations. (A and B) ER-phagy levels and quantitative analysis in the liver of ER-TRG (+/−) mice under ad libitum feeding and starvation conditions, respectively. Mice underwent a 16-h fast with free access to water, while control mice had ad libitum access to food and water. The pixel number of RFP-only, RFP⁺LAMP1⁺, and total area were calculated by ImageJ, and the ratio of RFP-only (or RFP⁺LAMP1⁺): the total area (RFP+GFP) was quantified. Values were stated as mean ± SEM (n = 5 per condition). Statistical differences were evaluated using Student’s t test. **P < 0.01, ***P < 0.001. Scale bar: 20 μm. (C and D) Quantitative analysis of ER-phagy levels in the liver of ER-TRG (+/−) mice with hepatic cancer. Confocal imaging shows reduced ER-phagy level at tumor sites compared to control liver or adjacent tissue. HC refers to hepatic cancer. Values were stated as mean ± SEM (n = 4 per condition). Statistical differences were assessed using Student’s t test. ***P < 0.001. Scale bar: 20 μm. (E and F) Representative confocal images and quantitative analysis of ER-phagy levels in the tibialis anterior (TA) muscle following skeletal muscle injury induced by 10 μM cardiotoxin compared with control tissues (the other leg in the same mouse). TA samples were collected and analyzed via confocal imaging on post-injury days 1, 3, and 7. Values were stated as mean ± SEM (n = 4 per condition). Statistical differences were determined using one-way ANOVA and Kruskal–Wallis test. *P < 0.05, **P < 0.01, *P < 0.001. Scale bar: 20 μm. See also Fig. S4.