Journal: Autophagy

Article Title: RTN4/Nogo-A-S1PR2 negatively regulates angiogenesis and secondary neural repair through enhancing vascular autophagy in the thalamus after cerebral cortical infarction.

doi: 10.1080/15548627.2022.2047344

Figure Lengend Snippet: Figure 2. Colocalization of RTN4 on different cell types in the ipsilateral thalamus after cortical infarction. (A) Triple-staining of RTN4 (blue) with MAP2+ neurons, CLDN11+ oligodendrocytes, GFAP+ astrocytes and AIF1+ microglia (green) and their spatial distribution along RECA-1+ vessels (red) within the ipsilateral thalamus at 7 days after MCAO (arrows). Insets show RTN4-expressing neurons and oligodendrocytes adjacent to RECA-1-labeled vessels. Scale bar: 50 μm. (B) Immunoblotting shows RTN4 expression in the ipsilateral thalamus of the sham- and MCAO-operated animals. (C) Quantitative analysis of RTN4 level relative to TUBA. n = 4, data are expressed as median ± interquartile range. *P < 0.05, compared with the sham-operated controls.

Article Snippet: Recombinant rat RTN4 Fc chimera corresponding to the specific amino acids 544–725 domain of RTN4 (R&D Systems, 2445-NG) was administered stereotactically into the right lateral ventricle to activate RTN4-Δ20-S1PR2 signaling [24].

Techniques: Staining, Expressing, Labeling, Western Blot

Journal: Autophagy

Article Title: RTN4/Nogo-A-S1PR2 negatively regulates angiogenesis and secondary neural repair through enhancing vascular autophagy in the thalamus after cerebral cortical infarction.

doi: 10.1080/15548627.2022.2047344

Figure Lengend Snippet: Figure 4. Knockdown of Rtn4 mediated by siRNA in the ipsilateral thalamus after cortical infarction. (A) Schematic diagram of brain section (−2.80 mm from bregma) shows the location of cortical infarction (black area) and the ipsilateral thalamus (blue area) where siRNA-lentivirus is delivered to four sites (red). (B) GFP-tagged Scramble- and Rtn4-siRNA were detected in the ipsilateral thalamus at 7 days after MCAO. Scale bar: 100 μm. (C) Immunostaining shows GFP-tagged Rtn4-siRNA to be expressed predominantly on MAP2+ neurons (arrows) and CLDN11+ oligodendrocytes (arrowheads). Scale bar: 50 μm. (D) Immunoblotting shows RTN4 expression in the ipsilateral thalamus from the Scramble- and Rtn4-siRNA groups at 7 days after MCAO. (E) Quantitative analysis of RTN4 level relative to TUBA. n = 4, data are expressed as median ± interquartile range. *P < 0.05, compared with the Scramble-siRNA group.

Article Snippet: Recombinant rat RTN4 Fc chimera corresponding to the specific amino acids 544–725 domain of RTN4 (R&D Systems, 2445-NG) was administered stereotactically into the right lateral ventricle to activate RTN4-Δ20-S1PR2 signaling [24].

Techniques: Knockdown, Immunostaining, Western Blot, Expressing

Journal: Autophagy

Article Title: RTN4/Nogo-A-S1PR2 negatively regulates angiogenesis and secondary neural repair through enhancing vascular autophagy in the thalamus after cerebral cortical infarction.

doi: 10.1080/15548627.2022.2047344

Figure Lengend Snippet: Figure 5. Rtn4 knockdown and vascular autophagic activation, as well as angiogenesis the ipsilateral thalamus after cortical infarction. (A) Immunoblotting shows the expression of S1PR2, RHOA and ROCK1 in vessels of the ipsilateral thalamus in the Scramble- and Rtn4-siRNA groups at 7 days after MCAO. (B-D) Quantitative analysis of S1PR2, RHOA and ROCK1 levels relative to TUBA. n = 4, data are expressed as median ± interquartile range. *P < 0.05, compared with the Scramble-siRNA group. (E) Immunoblotting shows the expression of MAP1LC3B-II, BECN1 and SQSTM1 in vessels of the ipsilateral thalamus in the Scramble- and Rtn4-siRNA groups. (F-H) Quantitative analysis of MAP1LC3B-II, BECN1 and SQSTM1 levels relative to GAPDH. n = 4, data are expressed as median ± interquartile range. *P < 0.05, compared with the Scramble-siRNA group. (I) Co-staining of BrdU (green) with LAMA1 (red) in the ipsilateral thalamus of the Scramble- and Rtn4-siRNA groups at 7 days after MCAO (arrows). Scale bar: 50 μm, 100 μm. (J and K) Quantitative analysis of BrdU+_LAMA1+ cells and vessel density. n = 6, data are expressed as median ± interquartile range. **P < 0.01, compared with the Scramble-siRNA group.

Article Snippet: Recombinant rat RTN4 Fc chimera corresponding to the specific amino acids 544–725 domain of RTN4 (R&D Systems, 2445-NG) was administered stereotactically into the right lateral ventricle to activate RTN4-Δ20-S1PR2 signaling [24].

Techniques: Knockdown, Activation Assay, Western Blot, Expressing, Staining

Journal: Autophagy

Article Title: RTN4/Nogo-A-S1PR2 negatively regulates angiogenesis and secondary neural repair through enhancing vascular autophagy in the thalamus after cerebral cortical infarction.

doi: 10.1080/15548627.2022.2047344

Figure Lengend Snippet: Figure 6. Activation of RTN4-S1PR2 and vascular autophagic activation, as well as angiogenesis in the ipsilateral thalamus after cortical infarction. (A) Immunoblotting shows the expression of S1PR2, RHOA, and ROCK1 in vessels of the ipsilateral thalamus in the IgG-Fc and RTN4-Δ20-Fc groups at 7 days after MCAO. (B-D) Quantitative analysis of S1PR2, RHOA and ROCK1 levels relative to TUBA. n = 4, data are expressed as median ± interquartile range. *P < 0.05, compared with the IgG-Fc group. (E) Immunoblotting shows the expression of MAP1LC3B-II, BECN1 and SQSTM1 in vessels of the ipsilateral thalamus in the IgG-Fc and RTN4- Δ20-Fc groups at 7 days after MCAO. (F-H) Quantitative analysis of MAP1LC3B-II, BECN1 and SQSTM1 levles relative to GAPDH. n = 4, data are expressed as median ± interquartile range. *P < 0.05, compared with the IgG-Fc group. (I) Co-staining for BrdU (green) and LAMA1 (red) within the ipsilateral thalamus in the IgG-Fc and RTN4-Δ20-Fc group at 7 days after MCAO (arrows). Scale bar: 50 μm, 100 μm. (J and K) Quantitative analysis of BrdU+_LAMA1+ cells and vessel density. n = 6, data are expressed as median ± interquartile range. **P < 0.01, compared with the IgG-Fc group.

Article Snippet: Recombinant rat RTN4 Fc chimera corresponding to the specific amino acids 544–725 domain of RTN4 (R&D Systems, 2445-NG) was administered stereotactically into the right lateral ventricle to activate RTN4-Δ20-S1PR2 signaling [24].

Techniques: Activation Assay, Western Blot, Expressing, Staining

Journal: Autophagy

Article Title: RTN4/Nogo-A-S1PR2 negatively regulates angiogenesis and secondary neural repair through enhancing vascular autophagy in the thalamus after cerebral cortical infarction.

doi: 10.1080/15548627.2022.2047344

Figure Lengend Snippet: Figure 10. Cognitive function and secondary neuronal damage of the ipsilateral thalamus after cortical infarction. (A) The representative swimming path of the animals in the Scramble-siRNA, Rtn4-siRNA, IgG-Fc, and RTN4-Δ20-Fc groups at 7 days after MCAO. (B) Animals in the Rtn4-siRNA group showed a shorter escape latency to reach the platform on day 6 after MCAO than that in the Scramble-siRNA group. In contrast, animals treated with RTN4-Δ20-Fc had longer escape latency from day 4–6 after MCAO than those in the IgG-Fc group. **P < 0.01, compared with the Scramble-siRNA group, ##P < 0.01, compared with the IgG-Fc group. (C) On day 7 after MCAO, animals of the Rtn4-siRNA group spent longer target quadrant time than that in the Scramble-siRNA group while animals treated with RTN4-Δ20-Fc showed less time in the target quadrant compared to the IgG-Fc group. n = 8, data are expressed as median ± interquartile range. *P < 0.05, compared with the Scramble-siRNA group, ##P < 0.01, compared with the IgG-Fc group. (D) Representative Nissl staining indicates the cortical infarction (black triangle) and neuronal damage within the ipsilateral thalamus (black rectangle) of the Scramble-siRNA, Rtn4-siRNA, IgG-Fc, and RTN4-Δ20-Fc groups. Immunostaining analysis of MAP2+

Article Snippet: Recombinant rat RTN4 Fc chimera corresponding to the specific amino acids 544–725 domain of RTN4 (R&D Systems, 2445-NG) was administered stereotactically into the right lateral ventricle to activate RTN4-Δ20-S1PR2 signaling [24].

Techniques: Staining, Immunostaining

Journal: Autophagy

Article Title: RTN4/Nogo-A-S1PR2 negatively regulates angiogenesis and secondary neural repair through enhancing vascular autophagy in the thalamus after cerebral cortical infarction.

doi: 10.1080/15548627.2022.2047344

Figure Lengend Snippet: Figure 11. RTN4-S1PR2 induced autophagy in BMVECs. (A) Immunoblotting analysis shows the expression of MAP1LC3B and SQSTM1 in BMVECs in the absence or presence of RTN4-Δ20 individual, or in combination with BafA1, 3-MA or JTE-013. (B and C) Quantitative analysis of MAP1LC3B-II and SQSTM1 levels relative to GAPDH. n = 4 from four independent experiments, data are expressed as median ± interquartile range. **P < 0.01, one-way ANOVA. (D) Representative confocal microscopic images of LC3B puncta in BMVECs transfected with mRFP-GFP-LC3 in the absence or presence of RTN4-Δ20 alone or in combination. Autophagosomes are indicated by yellow dots with both GFP and RFP, and autolysosomes by free red dots with only RFP in merged images. Scale bar: 20 μm. (E) Quantitative analysis of autophagosomes and autolysosomes per cell. n = 20–30 cells of four independent experiments, data are expressed as median ± interquartile range. **P < 0.01 (autophagosomes), ##P < 0.01 (autolysosomes), one-way ANOVA.

Article Snippet: Recombinant rat RTN4 Fc chimera corresponding to the specific amino acids 544–725 domain of RTN4 (R&D Systems, 2445-NG) was administered stereotactically into the right lateral ventricle to activate RTN4-Δ20-S1PR2 signaling [24].

Techniques: Western Blot, Expressing, Transfection

Journal: Advances in Radiation Oncology

Article Title: Kupffer Cell-Derived Interleukin-6 Aggravates Radiation-Induced Liver Disease by Activating Hepatocyte STAT3 to Promote Ccng1 Transcription

doi: 10.1016/j.adro.2026.102003

Figure Lengend Snippet: Schematic of the Kuppfer cell (KC)-hepatocyte crosstalk mechanism in RILD. Irradiation (IR) stimulates KCs to secrete IL-6, which binds to the IL-6R/gp130 complex on hepatocytes to activate JAK; phosphorylated JAK induces STAT3 phosphorylation, and nuclear-translocated p-STAT3 binds to the Ccng1 promoter to promote its transcription; CCNG1 then regulates MDM2 to mediate ubiquitination-dependent TP53 proteolysis, ultimately enhancing hepatocyte apoptosis and driving radiation-induced liver disease (RILD) progression.

Article Snippet: Either a rat interleukin (IL)-6 antibody (anti-IL-6; R&D Systems, #AF506) or soluble rat gp130 Fc chimera protein (sgp130Fc; R&D Systems, #5029-RG-100) was intraperitoneally injected twice weekly at concentrations of 16.7 μg/kg and 0.5 mg/kg, respectively.

Techniques: Irradiation, Phospho-proteomics, Ubiquitin Proteomics