Journal: World journal of gastroenterology

Article Title: Role of gut microbiota in Crohn's disease pathogenesis: Insights from fecal microbiota transplantation in mouse model.

doi: 10.3748/wjg.v30.i31.3689

Figure Lengend Snippet: Figure 5 Effects of gut microbiota from Crohn’s disease patients on the histopathological characteristics of mesenteric adipose and intestinal tissues from mice with 2,4,6-trinitrobenzene sulfonic acid-induced Crohn’s disease. A: Mesenteric adipose tissues collected from mice were stained with hematoxylin-eosin to assess histopathological alterations (× 150); B and C: Real-time reverse transcriptase-polymerase chain reaction assays of interleukin-6 (IL-6), IL-1β, tumour necrosis factor alpha, MCP-1, leptin, adiponectin, LXR, and FXR mRNA levels in mouse tissues; D: Intestinal tissues from mice stained with Masson stain to evaluate fibrotic alterations (× 150); E: Immunoblotting assays of alpha-smooth muscle actin and vimentin protein levels in mouse intestinal tissues. n = 6 or 3, bP < 0.01 vs control; aP < 0.05, bP < 0.01 vs 2,4,6-trinitrobenzene sulfonic acid (TNBS) alone; bP < 0.01 vs TNBS + NC-fetal microbiota transplantation, by one-way ANOVA with post-hoc Tukey HSD test. TNBS: 2,4,6-trinitrobenzene sulfonic acid; CD: Crohn’s disease; NC: Normal control; FMT: Fetal microbiota transplantation; IFN-γ: Interferon-gamma; TNF-α: Tumour necrosis factor alpha; IL: Interleukin; HE: Hematoxylin and eosin; α-SMA: Alpha-smooth muscle actin.

Article Snippet: The membranes were fixed for 1 hour using 5% nonfat milk in Trisbuffered saline with 0.1% Tween (TBS-T), rinsed three times with TBS-T, and incubated overnight at 4 °C with primary antibodies against α-SMA, vimentin, occluding, ZO-1, E-cadherin, cannabinoid receptor 1 (CB1), CB2, and GAPDH (dilution 1:1000, Proteintech, Wuhan, Hubei Province, China).

Techniques: Staining, Reverse Transcription, Polymerase Chain Reaction, Western Blot, Control, Transplantation Assay

Journal: International journal of biological sciences

Article Title: Contact Cooling-Induced ELOVL4 Enhances Skin Wound Healing by Promoting the Inflammation-to-Proliferation Phase Transition.

doi: 10.7150/ijbs.107871

Figure Lengend Snippet: Figure 1. 20°C treatment promotes skin wound healing. A. Schematic of wound healing rate in different seasons. B. K14 and VIM immunostaining shows 20°C treatment promotes wound healing. Scale bars, 50μm. Statistical of the length of the regenerated epidermis. N ≥ 5, **p < 0.01, ns: no significance. C. Volcano Plot shows different expressed genes in the 20°C-treatment group and control group. D. GO shows skin development pathway enriched in the 20°C-treatment group. E. KEGG shows fatty acid elongation pathway enriched in the 20°C-treatment group. F. iPATH shows fatty acid elongation pathway increased after 20°C treatment.

Article Snippet: The following antibodies were used in the study: K14 (Boster, BM4522), VIM (BEYOTIME, AF0318), ELOVL4 (Abclonal, A3639), PCNA (Elabscience, E-AB-22001), CD31 (Servicebio, GB12063-100), TNFα (BEYOTIME, AF8208), P63 (Zenbio, 381215), IL1β (BEYOTIME, AF7209).

Techniques: Immunostaining, Control

Journal: International journal of biological sciences

Article Title: Contact Cooling-Induced ELOVL4 Enhances Skin Wound Healing by Promoting the Inflammation-to-Proliferation Phase Transition.

doi: 10.7150/ijbs.107871

Figure Lengend Snippet: Figure 3. Elovl4-EPA/DHA promotes wound healing. A. Phase-contrast microscope and statistics show wound healing after long chain fatty acid treatment; Statistics of average wound size. Scale bars, 2 mm. N ≥ 5, ***p < 0.001, **p < 0.01, *p < 0.05. B. K14/PCNA and K14/CD31 immunostaining shows wound healing after DHA, EPA, or ELOVL4 inhibitor treatment; Statistics of average PCNA+ and CD31+ cells. Scale bars, 200μm. N ≥ 5, **p < 0.01, *p < 0.05. C. Schematic shows wound healing after DHA, EPA, or ELOVL4 inhibitor treatment.

Article Snippet: The following antibodies were used in the study: K14 (Boster, BM4522), VIM (BEYOTIME, AF0318), ELOVL4 (Abclonal, A3639), PCNA (Elabscience, E-AB-22001), CD31 (Servicebio, GB12063-100), TNFα (BEYOTIME, AF8208), P63 (Zenbio, 381215), IL1β (BEYOTIME, AF7209).

Techniques: Microscopy, Immunostaining

Journal: International journal of biological sciences

Article Title: Contact Cooling-Induced ELOVL4 Enhances Skin Wound Healing by Promoting the Inflammation-to-Proliferation Phase Transition.

doi: 10.7150/ijbs.107871

Figure Lengend Snippet: Figure 5. 20°C treatment can inhibit TNFα-mediated inflammatory response. A. FeaturePlots show increased expression of Il1b and Tnfa in PWD3. B. qRT-PCR shows the expression of inflammatory factors decreased in the 20°C group. N ≥ 5, **p < 0.01, ***p < 0.001, ns: no significance. C. TNFα immunostaining shows the decreased expression of TNFα in the 20°C-treatment group; Statistics of average FI of TNFα. Scale bars, 100μm. N ≥ 5, **p < 0.01, *p < 0.05. D. K14/VIM, K14/CD31, K14/PCNA, and P63/VIM immunostaining shows wound healing in the TNFα group and the 20°C treatment +TNFα group. E. Statistics of the average rate of re-epithelialization. Scale bars, 100μm. N ≥ 5, **p < 0.01, *p < 0.05. F. Schematic shows decreased expression of TNFα after 20°C treatment.

Article Snippet: The following antibodies were used in the study: K14 (Boster, BM4522), VIM (BEYOTIME, AF0318), ELOVL4 (Abclonal, A3639), PCNA (Elabscience, E-AB-22001), CD31 (Servicebio, GB12063-100), TNFα (BEYOTIME, AF8208), P63 (Zenbio, 381215), IL1β (BEYOTIME, AF7209).

Techniques: Expressing, Quantitative RT-PCR, Immunostaining

Journal: International journal of biological sciences

Article Title: Contact Cooling-Induced ELOVL4 Enhances Skin Wound Healing by Promoting the Inflammation-to-Proliferation Phase Transition.

doi: 10.7150/ijbs.107871

Figure Lengend Snippet: Figure 6. Elovl4-EPA/DHA promotes skin organoids to regenerate skin. A. VlnPlot shows expression of Elovl4. B. FeaturePlot shows Elovl4 expressed in epidermal cells. C. Immunostaining shows that ELOVL4 is expressed in epidermal cells. D. TNFα/VIM and IL1β/VIM immunostaining shows the decreased expression of inflammatory factors after LCFAs treatment; Statistics of average FI of TNFα- and IL1β-expressing cells. Scale bars, 20μm. N ≥ 5, **p < 0.01, *p < 0.05, ns: no significance. E. Phase-contrast microscope and statistics show wound healing after long chain fatty acid treatment; Statistics of average wound size. N ≥ 5, **p < 0.01, *p < 0.05. F. K14 / PCNA and TNFα immunostaining shows depressed wound healing in the iELOVL4 transplantation group. Statistics of PCNA+ cells and average FI of TNFα. Scale bars, 100μm. N ≥ 5, *p < 0.05.

Article Snippet: The following antibodies were used in the study: K14 (Boster, BM4522), VIM (BEYOTIME, AF0318), ELOVL4 (Abclonal, A3639), PCNA (Elabscience, E-AB-22001), CD31 (Servicebio, GB12063-100), TNFα (BEYOTIME, AF8208), P63 (Zenbio, 381215), IL1β (BEYOTIME, AF7209).

Techniques: Expressing, Immunostaining, Microscopy, Transplantation Assay

Journal: PLoS ONE

Article Title: Hypoxia-Inducible Factor-1alpha and MAPK Co-Regulate Activation of Hepatic Stellate Cells upon Hypoxia Stimulation

doi: 10.1371/journal.pone.0074051

Figure Lengend Snippet: HSC-T6 cells were cultured in room air or in 1% oxygen. (A) Vimentin expression in HSC-T6 cells grown on the coverslips at 48 hours post-seeding were co-stained with anti-vimentin (red) and DAPI (blue) by immunocytochemistry. (B) Cells were collected at indicated time and cell lysates were subjected to detect vimentin and α-SMA. Densitometric analysis was performed using pooled data from three such experiments. Data are mean ± SD. (C) vimentin; (D) α-SMA. * : P <0.05.

Article Snippet: SDS-PAGE and western blot were performed to determine the expression of Hif-1α (2015-1, Epitomics, USA), vimentin (bs-0756R, Bioss, China), α-SMA (A7607, Sigma, USA), phosphorylated MAPK (2219-1, Epitomics, USA), GAPDH (KC-5G5, Kangchen, China), H2AFX (10856-1-AP, Proteintech, USA) and β-actin (sc-47778, Santa-Cruz, USA).

Techniques: Cell Culture, Expressing, Staining, Immunocytochemistry

Journal: PLoS ONE

Article Title: Hypoxia-Inducible Factor-1alpha and MAPK Co-Regulate Activation of Hepatic Stellate Cells upon Hypoxia Stimulation

doi: 10.1371/journal.pone.0074051

Figure Lengend Snippet: HSC-T6 cells in 6-well plates were transfected with either 100 nM Hif-1α siRNA or nonspecific (NS) siRNA for 24h and then cultured in room air or in 1% oxygen for 48h. (A) Cells were collected and cell lysates were subjected to detect Hif-1α, vimentin, α-SMA and β-actin with western blot; Densitometric analysis was performed using pooled data from three such experiments. Data are mean ± SD. (B) Hif-1α; (C) vimentin; (D) α-SMA. * : P <0.05, ** : P <0.01 (RA: room air, KD: knock down)..

Article Snippet: SDS-PAGE and western blot were performed to determine the expression of Hif-1α (2015-1, Epitomics, USA), vimentin (bs-0756R, Bioss, China), α-SMA (A7607, Sigma, USA), phosphorylated MAPK (2219-1, Epitomics, USA), GAPDH (KC-5G5, Kangchen, China), H2AFX (10856-1-AP, Proteintech, USA) and β-actin (sc-47778, Santa-Cruz, USA).

Techniques: Transfection, Cell Culture, Western Blot