Journal: Cellular & molecular biology letters

Article Title: MAFF alleviates hepatic ischemia-reperfusion injury by regulating the CLCF1/STAT3 signaling pathway.

doi: 10.1186/s11658-025-00721-x

Figure Lengend Snippet: Fig. 4 MAFF relieves inflammation in hepatic IRI. A At 6 h after hepatic IRI, the liver tissues of both rAd-NC and rAd-shMAFF mice were analyzed to assess mRNA levels of inflammatory cytokines and chemokines (n = 3). B and C Immunofluorescence was performed to detect CD11b-positive inflammatory cells (red) in ischemic liver sections obtained from mice in the specified experimental groups (n = 6); scale bar = 50 and 100 μm. D and E IHC was performed to assess Ly6G expression in ischemic liver sections obtained from mice in the specified experimental groups (n = 6); scale bar = 100 μm. F At 6 h after hepatic IRI, the liver tissues of both the rAd-NC and the rAd-OEMAFF mice were analyzed to assess mRNA levels of inflammatory cytokines and chemokines (n = 3). G and H Immunofluorescence staining was used to identify inflammatory cells expressing CD11b (red) in liver sections from the mice assigned to the designated experimental groups (n = 6); scale bars = 50 and 100 μm. I and J IHC was conducted to evaluate the presence of Ly6G in liver sections affected by ischemia, using samples obtained from mice assigned to specific experimental groups (n = 6); scale bar = 100 μm

Article Snippet: The sections were treated with MAFF (12,771–1-AP, Proteintech, 1:100), Ly6g (ab303467, Abcam, 1:100), and P-STAT3 (ab76315, Abcam, 1:100) at 4 °C overnight.

Techniques: Immunofluorescence, Expressing, Staining

Journal: Cellular & molecular biology letters

Article Title: MAFF alleviates hepatic ischemia-reperfusion injury by regulating the CLCF1/STAT3 signaling pathway.

doi: 10.1186/s11658-025-00721-x

Figure Lengend Snippet: Fig. 6 Hepatocyte apoptosis is regulated by the MAFF-mediated CLCF1 pathway. A–B Flow cytometry was used to evaluate cell apoptosis in hepatocytes exposed to shCLCF1 (n = 3). C–D The impact of CLCF1 on AML12 cell apoptosis both with and without MAFF knockdown, as determined using flow cytometry (n = 3). E WB was used to assess STAT3 activation in AML12 cells subjected to either normoxia or H/R in the shNC and shMAFF groups. F WB was used to assess STAT3 activation in AML12 cells subjected to either normoxia or H/R in the shNC group and shCLCF1 group. G WB was used to assess STAT3 activation in AML12 cells under H/R stimulation following the addition of exogenous CLCF1 in the shNC and shMAFF groups

Article Snippet: The sections were treated with MAFF (12,771–1-AP, Proteintech, 1:100), Ly6g (ab303467, Abcam, 1:100), and P-STAT3 (ab76315, Abcam, 1:100) at 4 °C overnight.

Techniques: Flow Cytometry, Knockdown, Activation Assay

Journal: Cellular & molecular biology letters

Article Title: MAFF alleviates hepatic ischemia-reperfusion injury by regulating the CLCF1/STAT3 signaling pathway.

doi: 10.1186/s11658-025-00721-x

Figure Lengend Snippet: Fig. 7 MAFF binding to BACH1 facilitates the activation of the CLCF1/STAT3 signaling pathway. A–E Following immunoprecipitation (IP) with MAFF antibody or IgG (negative control antibody), samples collected from hepatocytes were subjected to electrophoresis. Subsequently, Coomassie brilliant blue staining was performed. This staining technique was employed after the samples were subjected to H/R. The present study employed mass spectrum analysis to examine IP-MAFF and IP-IgG lysates. A Venn diagram was generated, and GO analysis was performed. F Interactions among 50 proteins. G Co-IP of MAFF and BACH1 in hepatocytes after H/R. H Immunofluorescence staining of MAFF, BACH1, and DAPI in hepatocytes after H/R; scale bar = 20 μm. I RT-qPCR was used to assess CLCF1 levels (n = 3). J The effect of BACH1 on the STAT3 pathway in liver cells was evaluated using WB analysis after suppressing BACH1 expression. K A luciferase test was conducted utilizing the pGL4-luciferase vector. The transcriptional activity of the discovered MARE was reduced upon BACH1 or MAFF knockdown (n = 3)

Article Snippet: The sections were treated with MAFF (12,771–1-AP, Proteintech, 1:100), Ly6g (ab303467, Abcam, 1:100), and P-STAT3 (ab76315, Abcam, 1:100) at 4 °C overnight.

Techniques: Binding Assay, Activation Assay, Immunoprecipitation, Negative Control, Electrophoresis, Staining, Generated, Co-Immunoprecipitation Assay, Immunofluorescence, Quantitative RT-PCR, Expressing, Luciferase, Plasmid Preparation, Activity Assay, Knockdown

Journal: Cellular & molecular biology letters

Article Title: MAFF alleviates hepatic ischemia-reperfusion injury by regulating the CLCF1/STAT3 signaling pathway.

doi: 10.1186/s11658-025-00721-x

Figure Lengend Snippet: Fig. 8 Liver IRI is protected in vivo by MAFF via the CLCF1/STAT3 pathway. A Hepatic IRI model. B and F Histological analysis using H&E staining revealed the presence of necrotic areas in the liver tissue (n = 5); scale bars = 100 μm and 20 μm. C and G IHC was conducted to evaluate P-STAT3 expression in liver sections (n = 5); scale bars = 100 μm and 20 μm. D and H TUNEL staining and statistical analysis were performed on ischemic liver sections obtained from the mice; scale bar = 50 μm. E and I Immunofluorescence staining was used to identify inflammatory cells expressing CD11b (red) in liver slices affected by ischemia (n = 5); scale bar = 100 μm. J and K ALT and AST levels in the blood were assessed in the rAd-NC, rAd-shMAFF, and rAd-shMAFF + CLCF1 groups (n = 5)

Article Snippet: The sections were treated with MAFF (12,771–1-AP, Proteintech, 1:100), Ly6g (ab303467, Abcam, 1:100), and P-STAT3 (ab76315, Abcam, 1:100) at 4 °C overnight.

Techniques: In Vivo, Staining, Expressing, TUNEL Assay, Immunofluorescence