Journal: Stem Cell Research & Therapy

Article Title: The adipose-derived mesenchymal stem cell secretome promotes hepatic regeneration in miniature pigs after liver ischaemia-reperfusion combined with partial resection

doi: 10.1186/s13287-021-02284-y

Figure Lengend Snippet: Effects of ASCs and the ASC-secretome on liver regeneration-related proteins. a Immunohistochemical staining of the Ki67 protein in liver tissue sections at all time points in the four groups. b Results of Ki67 immunohistochemical analysis. c Western blot of liver regeneration-related proteins. d Analysis of PCNA protein expression levels. Figure 4e: Analysis of p-STAT3 protein expression levels. f Analysis of SOCS3 protein expression levels. The data are expressed as the mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, versus the saline group. # p < 0.05, ## p < 0.01, versus the DMEM group.

Article Snippet: The membrane was blocked in TBST containing 5% nonfat powdered milk for 2 h at room temperature to prevent nonspecific binding, incubated with primary antibody overnight at 4 °C, and then incubated with a horseradish peroxidase (HRP)-conjugated anti-species secondary antibody for 2 h. Antibodies against PCNA (Abcam, Cambridge, UK), p-STAT3, STAT3, suppressor of cytokine signalling 3 (SOCS3) (ImmunoWay, Plano, USA) and β-actin and a horseradish peroxidase (HRP)-conjugated antibody (Sangon Biotech, Shanghai, China) were used.

Techniques: Immunohistochemical staining, Staining, Western Blot, Expressing

Journal: Stem Cell Research & Therapy

Article Title: The adipose-derived mesenchymal stem cell secretome promotes hepatic regeneration in miniature pigs after liver ischaemia-reperfusion combined with partial resection

doi: 10.1186/s13287-021-02284-y

Figure Lengend Snippet: The experimental procedure and the mechanism by which the ASC-secretome promotes liver regeneration. The model of liver ischaemia-reperfusion combined with partial hepatectomy was established by laparoscopy. The ASC-secretome of miniature pigs was prepared and injected into the liver parenchyma immediately after the operation. The ASC-secretome exerted obvious anti-inflammatory and liver regenerative effects. The mechanism may be as shown in Fig. 5. After partial hepatectomy, the STAT3 signalling pathway was activated to promote the proliferation of hepatocytes. Furthermore, a SOCS3 negative feedback loop was activated to terminate the STAT3 signalling cascade. The ASC-secretome acted on SOCS3 to reduce its expression and interrupted the negative feedback regulation of the STAT3 signalling pathway, accelerating the process of liver regeneration. Cytokines in the ASC-secretome regulate inflammation and promote liver regeneration.

Article Snippet: The membrane was blocked in TBST containing 5% nonfat powdered milk for 2 h at room temperature to prevent nonspecific binding, incubated with primary antibody overnight at 4 °C, and then incubated with a horseradish peroxidase (HRP)-conjugated anti-species secondary antibody for 2 h. Antibodies against PCNA (Abcam, Cambridge, UK), p-STAT3, STAT3, suppressor of cytokine signalling 3 (SOCS3) (ImmunoWay, Plano, USA) and β-actin and a horseradish peroxidase (HRP)-conjugated antibody (Sangon Biotech, Shanghai, China) were used.

Techniques: Injection, Expressing

Journal: bioRxiv

Article Title: Retinal ganglion cell survival after severe optic nerve injury is modulated by crosstalk between JAK/STAT signaling and innate immune responses in the zebrafish retina

doi: 10.1101/2021.04.08.439090

Figure Lengend Snippet: (A) pSTAT3 expression (red) in flat-mount isl2b :GFP ONT- and ONT+ retinae at 1dpi. Nuclei stained with DAPI (blue). Boxed regions are 3x zooms of 40x images. (B) Quantification of pSTAT3 levels at 1dpi. pSTAT3 levels in ONT+ RGCs relative to levels normalized to those in ONT- RGCs. Shown are mean±SD of n=3 for each group; **p<0.01; Mann-Whitney test. (C) Experimental paradigm to assess Jak requirement during RGC survival after ONT. (D) pSTAT3 expression in ONT+ isl2b :GFP retinal flat mounts at 1 and 7dpi +/-intravitreal (IV) injection of the Jak inhibitor, P6. DMSO was used as control (Ctr). (E) Quantification of pSTAT3 expression after P6 application at 1 and 7dpi. N=3/condition. Shown are mean±SD; *p<0.05, Mann-Whitney test. (F) Images of 7dpi P6- or DMSO-injected flat-mount isl2b :GFP retinae. (G) Quantification of RGC survival after P6 injection (n=5/condition). Shown are mean±SD; **p<0.01; Kruskal Wallis ANOVA with Dunn’s multiple comparisons. Scale bars = 50μm and 150μm on zoomed images in A.

Article Snippet: Immunofluorescence staining on retinal cryosections and flat-mounted retinae were performed as previously described ( ; ) with the addition of an antigen retrieval step consisting of 100% methanol incubation at −20 C for 30 minutes for staining pSTAT3 (MBL International Corporation, D128-3).

Techniques: Expressing, Staining, MANN-WHITNEY, IV Injection, Injection