Journal: PLOS ONE

Article Title: High glucose promotes macrophage switching to the M1 phenotype via the downregulation of STAT-3 mediated autophagy

doi: 10.1371/journal.pone.0314974

Figure Lengend Snippet: Double-immunofluorescence images and quantification of CD68 and iNOS (A and B), CD68 and MR (C and D), CD68 and Arg-1 (E and F), CD68 and p-STAT-3 (G and H), CD68 and LC3 (I and J) were detection in rat kidney tissue (white arrows) (×400; bars = 50μm). MR: Mannose Receptor; Arg-1: Arginase 1.

Article Snippet: Then slides were incubated with primary antibodies: anti-CD68 antibody (Santa Cruz, sc-70761), anti-iNOS antibody (Abcam, ab15323), anti-MR antibody (Abcam, ab64693), anti-Arg-1 antibody (Sigma, ABS535), anti-p-STAT-3-Y705 antibody (Sigma, SAB5700314), anti-LC3 antibody (Sigma, L7543), CD11c (Santa Cruz, sc-398708), CD206 (Santa Cruz, sc-70585), respectively, overnight at 4°C.

Techniques: Immunofluorescence

Journal: PLOS ONE

Article Title: High glucose promotes macrophage switching to the M1 phenotype via the downregulation of STAT-3 mediated autophagy

doi: 10.1371/journal.pone.0314974

Figure Lengend Snippet: A-F, Representative Western blotting analysis and quantification of LC3, Beclin-1 (C), iNOS, MR, p-STAT-3. GAPDH was used as an internal control. Data are presented as mean± SD (n = 3). *P<0.05 vs NC (normal control), respectively. G, the immunofluorescence analysis of HG (high glucose) on macrophage phenotype (×400; bars = 20μm). NC:11mmol/L glucose, HG: 30 mmol/L glucose; Mannitol: 11mmol/L glucose + 19 mmol/L mannitol. MR: Mannose Receptor.

Article Snippet: Then slides were incubated with primary antibodies: anti-CD68 antibody (Santa Cruz, sc-70761), anti-iNOS antibody (Abcam, ab15323), anti-MR antibody (Abcam, ab64693), anti-Arg-1 antibody (Sigma, ABS535), anti-p-STAT-3-Y705 antibody (Sigma, SAB5700314), anti-LC3 antibody (Sigma, L7543), CD11c (Santa Cruz, sc-398708), CD206 (Santa Cruz, sc-70585), respectively, overnight at 4°C.

Techniques: Western Blot, Control, Immunofluorescence

Journal: PLOS ONE

Article Title: High glucose promotes macrophage switching to the M1 phenotype via the downregulation of STAT-3 mediated autophagy

doi: 10.1371/journal.pone.0314974

Figure Lengend Snippet: L, Representative Western blotting analysis and quantification (B) of LC3, Beclin-1, iNOS, MR, p-STAT-3. GAPDH was used as an internal control. Data are presented as mean ± SD (n = 3). M, the immunofluorescence analysis of STAT-3 activator and autophagy activator (Rapamycin, Rapa) on macrophage phenotype (×400; bars = 20μm). *P<0.05 vs NC (normal control), # P<0.05 vs HG. NC:11mmol/L glucose, HG: 30 mmol/L glucose; Mannitol: 11mmol/L glucose + 19 mmol/L mannitol. MR: Mannose Receptor; SA: STAT-3 activator, Rapa: Rapamycin.

Article Snippet: Then slides were incubated with primary antibodies: anti-CD68 antibody (Santa Cruz, sc-70761), anti-iNOS antibody (Abcam, ab15323), anti-MR antibody (Abcam, ab64693), anti-Arg-1 antibody (Sigma, ABS535), anti-p-STAT-3-Y705 antibody (Sigma, SAB5700314), anti-LC3 antibody (Sigma, L7543), CD11c (Santa Cruz, sc-398708), CD206 (Santa Cruz, sc-70585), respectively, overnight at 4°C.

Techniques: Western Blot, Control, Immunofluorescence

Journal: PLOS ONE

Article Title: High glucose promotes macrophage switching to the M1 phenotype via the downregulation of STAT-3 mediated autophagy

doi: 10.1371/journal.pone.0314974

Figure Lengend Snippet: A and B, The effect of STAT-3 siRNA on STAT-3 expression. *p < 0.05 vs. NC group; NC: Normal control; siRNA: Small interfering RNA; NTC: Nontarget control (NTC) siRNA. C-G: RAW264.7 cells were treated with different reagents. The cells were collected for western blot analysis, quantification analysis of p-STAT-3, LC3, iNOS and MR. data are presented as the mean ± SD (n = 3 per group). *p < 0.05 vs. NC group; #p < 0.05 vs. HG group, &p < 0.05 vs. HG+SA group, $p < 0.05 vs. HG+Rapa group. HG: High glucose; MR: Mannose receptor; STAT: Signal transducer and activator of transcription; Rapa: Rapamycin; SA: STAT-3 activator; 3-MA:3-Methyladenine. NC:11mmol/L glucose, HG: 30 mmol/L glucose; Mannitol: 11mmol/L glucose + 19 mmol/L mannitol.

Article Snippet: Then slides were incubated with primary antibodies: anti-CD68 antibody (Santa Cruz, sc-70761), anti-iNOS antibody (Abcam, ab15323), anti-MR antibody (Abcam, ab64693), anti-Arg-1 antibody (Sigma, ABS535), anti-p-STAT-3-Y705 antibody (Sigma, SAB5700314), anti-LC3 antibody (Sigma, L7543), CD11c (Santa Cruz, sc-398708), CD206 (Santa Cruz, sc-70585), respectively, overnight at 4°C.

Techniques: Expressing, Control, Small Interfering RNA, Western Blot