Journal: Kidney Diseases

Article Title: Treatment of Membranous Nephropathy by Disulfiram through Inhibition of Podocyte Pyroptosis

doi: 10.1159/000524164

Figure Lengend Snippet: DSF alleviated C3a/C5a-induced podocyte injury by inhibiting pyroptosis. Podocyte injury was induced with C3a (50 nM) and C5a (50 nM). The inhibitory effects of DSF (250 nM) on C3a/C5a-induced pyroptosis were examined. a Representative images of GSDMD(N)/ZO-1/Nucleus (DAPI) triple immunofluorescent staining of treated podocytes. Scale bars = 20 μm. b Representative Western Blot images of GSDMD, NF-κB p65, p-NF-κB p65 (Ser536), NLRP3, ASC, Caspase-1, IL-18 and the internal control (GAPDH) in treated podocytes. c IL-18 release in treated podocytes was detected. d, e Representative images of PI/Nucleus (DAPI) double fluorescent staining of treated podocytes ( d ) and percentage of PI-positive cells ( e ); arrows, PI-positive cells; scale bars = 40 μm. f LDH release in treated podocytes was detected. The data above represent three independent experiments in duplicate and are shown as the mean ± SD, and ANOVA with LSD-t test (equal variances assumed) or Welch's test with Dunnett's T3 test (equal variances not assumed) was used for multiple comparisons among groups. *, p < 0.05; **, p < 0.01.

Article Snippet: IL-18 release in the culture supernatant of the podocytes and the serum IL-1β/IL-18 levels of rats were tested using human IL-18 ELISA and rat IL-1β/IL-18 ELISA kits (Boster Biological Technology Co. Ltd., Wuhan, China), respectively.

Techniques: Staining, Western Blot

Journal: Kidney Diseases

Article Title: Treatment of Membranous Nephropathy by Disulfiram through Inhibition of Podocyte Pyroptosis

doi: 10.1159/000524164

Figure Lengend Snippet: DSF inhibited the renal pyroptosis signaling pathway in PHN rats. a Representative renal GSDMD(N)/Synaptopodin and GSDMD(N)/ZO-1 double immunofluorescent staining of the rats ( n = 6) in each group; Scale bars = 20 μm. b, c Representative renal immunohistochemical staining ( b ) and semiquantification based on the glomerular IOD/area of GSDMD(N), NF-κB p65, p-NF-κB p65 (Ser536), NLRP3, ASC, Caspase-1, IL-1β, and IL-18 ( c ) of the rats ( n = 6) in each group; Scale bars = 20 μm. d Relative mRNA levels of glomerular GSDMD, NLRP3, ASC, Caspase-1, IL-1β and IL-18 of the rats ( n = 6) in each group. e Representative Western Blot images of renal GSDMD, NF-κB p65, p-NF-κB p65 (Ser536), nuclear NF-κB p65, nuclear p-NF-κB p65 (Ser536), NLRP3, ASC, Caspase-1, Caspase-1 p20, IL-1β, IL-1β (mature form), IL-18 and the internal control (GAPDH, Histone H3) of the rats ( n = 6) in each group. Serum IL-1β ( f ) and IL-18 ( g ) of the rats ( n = 6) in each group on days 1, 5, 8, 15 after model establishment. The data above are shown as the mean ± SD ( c, d, f, g ) and were compared to the PHN group ( f, g ). ANOVA with LSD-t test (equal variances assumed) or Welch's test with Dunnett's T3 test (equal variances not assumed) was used for multiple comparisons among groups. *, p < 0.05; **, p < 0.01.

Article Snippet: IL-18 release in the culture supernatant of the podocytes and the serum IL-1β/IL-18 levels of rats were tested using human IL-18 ELISA and rat IL-1β/IL-18 ELISA kits (Boster Biological Technology Co. Ltd., Wuhan, China), respectively.

Techniques: Staining, Immunohistochemical staining, Western Blot

Journal: Cellular microbiology

Article Title: Adherence and internalization of Streptococcus gordonii by epithelial cells involves beta1 integrin recognition by SspA and SspB (antigen I/II family) polypeptides.

doi: 10.1111/j.1462-5822.2006.00768.x

Figure Lengend Snippet: Fig. 7. Expression of interleukin-8 by HEp-2 cells stimulated with streptococci. A. HEp-2 monolayers were grown to confluence in 12-well plates and infected with 108 cells of S. gordonii DL1 () or S. pyogenes A40 () for 90 min. The monolayers were then washed and incubated for up to 24 h in the presence of penicillin G (5 mg ml-1) and gentamycin (0.1 mg ml-1). Well contents, together with those of uninfected controls, were sampled at various times and IL-8 accumulation was determined by ELISA as described in Experimental procedures. HEp-2 monolayers were also infected with 108 cells of S. gordonii DL1 continuously for 24 h in the absence of antibiotics (hatched columns). Data shown are for 12 h and 24 h samples. Error bars are SD of the mean of two independent determinations performed in triplicate. Recombinant human interleukin-1a (10 ng ml-1) stimulation (positive control) resulted in 0.23 0.03 ng ml-1 IL-8 released by ELISA. B. HEp-2 monolayers were inoculated with 108 cells of S. gordonii DL1 or S. pyogenes A40, together with uninfected controls, and incubated for up to 12 h. At various intervals, RNA was extracted from cells and subjected to Northern blot analysis (10 mg of RNA per lane) with an IL-8 DNA probe. Lane 1, unstimulated control 1 h; lane 2, control 12 h; lane 3, S. gordonii DL1 1 h; lane 4, S. gordonii DL1 12 h; lane 5, S. pyogenes A40 1 h; lane 6, S. pyogenes A40 12 h. Visible degeneration of S. pyogenes-infected HEp-2 monolayers began to occur after 12 h.

Article Snippet: Levels of IL-8 released were determined using a human IL-8 DuoSet ELISA Development kit (R&D Systems).

Techniques: Expressing, Infection, Incubation, Enzyme-linked Immunosorbent Assay, Recombinant, Positive Control, Northern Blot, Control