rabbit anti human sod2  (Proteintech)

Journal: Animal Cells and Systems

Article Title: Epinephrine as a potential driver of oral lichen planus pathogenesis

doi: 10.1080/19768354.2025.2588914

Figure Lengend Snippet: High level of epinephrine induces intracellular oxidative stress in oral keratinocytes. (A) HOK-16B cells treated with PBS or epinephrine for 24 h were subjected to CellROX staining. Nuclear DAPI signal is shown in blue (left). The relative CellROX signal intensities are shown (right). Each symbol means an individual cell. (B) Immunoblot analysis of SOD2 and SESN2 in HOK-16B cells treated with epinephrine for 24 h. β ACTIN was used as loading control. (C) Relative band intensities are shown. (D) Relative mRNA expression of TNFα, IL6, and IL8 in HOK-16B cells treated with epinephrine for 3 h. ** P < 0.01; *** P < 0.001. n.s , not significant.

Article Snippet: After blocking with 5% non-fat dried milk in Tris-buffered saline with TweenTM 20 (TBST) (10 mM Tris, pH 8.0, 150 mM NaCl and 0.5% Tween 20) for 30 min, membranes were incubated with antibodies against phospho STAT3 (1:1000, Cell signaling, Danvers, MA, USA, cat no. 9145), total STAT3 (1:1000, Cell signaling, cat no. 8768), β ACTIN (1:5000, Sigma Aldrich, cat no. A5316), phospho ERK (1:1000, Thermo Fisher Scientific, Waltham, MA, USA, cat no. MA5-15174), total ERK (1:1000, Cell signaling, cat no. 4695), phospho AKT (1:1000, Cell signaling, cat no. 4058), total AKT (1:1000, Cell signaling, cat no. 9272), BCL2 (1:1000, Bioworld Technology, China, cat no. BS1511), SOD2 (1:1000, Cusabio, Houston, TX, USA, cat no. CSB-PA022398LA01HU), and SESN2 (1:1000, Abcam, Cambridge, UK, cat no. ab178518) overnight at 4°C.

Techniques: Staining, Western Blot, Control, Expressing

Journal: Frontiers in Immunology

Article Title: Single-cell sequencing analysis reveals cancer-associated pericyte subgroup in esophageal squamous cell carcinoma to predict prognosis

doi: 10.3389/fimmu.2024.1474673

Figure Lengend Snippet: Multiple fluorescence staining of tissues samples from ESCC patients. (A, B) Validation of high and low expression level of c5_TYMS in ESCC tumor tissues samples and spatial distribution with microphages and fibroblasts. (C, D) Validation of high and low expression of level c4_SOD2 in ESCC tumor tissue samples and spatial distribution with microphages and fibroblasts.

Article Snippet: Incubate overnight at 4°C with purified rabbit anti-human PDGFRβ (ab32570, 1:100, Abcam, USA), rabbit anti-human SOD2 (66474-1-Ig, 1:300, Proteintech, Wuhan, China), rabbit anti-human CD68 (66231-2-Ig, 1:2000, Proteintech, Wuhan, China), rabbit anti-human TYMS (66725-1-Ig, 1:200, Proteintech, Wuhan, China), mouse anti-human α-SMA(67735-1-Ig, 1:400, Proteintech, Wuhan, China) and purified rabbit anti-human EPCAM (21050-1-AP, 1:1000, Proteintech, Wuhan, China).

Techniques: Fluorescence, Staining, Biomarker Discovery, Expressing

Journal: Frontiers in Immunology

Article Title: Single-cell sequencing analysis reveals cancer-associated pericyte subgroup in esophageal squamous cell carcinoma to predict prognosis

doi: 10.3389/fimmu.2024.1474673

Figure Lengend Snippet: Docking results of available proteins with small molecules.

Article Snippet: Incubate overnight at 4°C with purified rabbit anti-human PDGFRβ (ab32570, 1:100, Abcam, USA), rabbit anti-human SOD2 (66474-1-Ig, 1:300, Proteintech, Wuhan, China), rabbit anti-human CD68 (66231-2-Ig, 1:2000, Proteintech, Wuhan, China), rabbit anti-human TYMS (66725-1-Ig, 1:200, Proteintech, Wuhan, China), mouse anti-human α-SMA(67735-1-Ig, 1:400, Proteintech, Wuhan, China) and purified rabbit anti-human EPCAM (21050-1-AP, 1:1000, Proteintech, Wuhan, China).

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: Single-cell sequencing analysis reveals cancer-associated pericyte subgroup in esophageal squamous cell carcinoma to predict prognosis

doi: 10.3389/fimmu.2024.1474673

Figure Lengend Snippet: Molecular docking results of predicted drugs and protein encoded by target genes. (A) PDGFRβ and docetaxel. (B) TYMS and docetaxel. (C) PDGFRβ and cytarabine. (D) TYMS and cytarabine. (E) PDGFRβ and bisindolylmalemide. (F) SOD2 and bisindolylma-leimide. (G) PDGFRβ and raloxifene. (H) SOD2 and raloxifene.

Article Snippet: Incubate overnight at 4°C with purified rabbit anti-human PDGFRβ (ab32570, 1:100, Abcam, USA), rabbit anti-human SOD2 (66474-1-Ig, 1:300, Proteintech, Wuhan, China), rabbit anti-human CD68 (66231-2-Ig, 1:2000, Proteintech, Wuhan, China), rabbit anti-human TYMS (66725-1-Ig, 1:200, Proteintech, Wuhan, China), mouse anti-human α-SMA(67735-1-Ig, 1:400, Proteintech, Wuhan, China) and purified rabbit anti-human EPCAM (21050-1-AP, 1:1000, Proteintech, Wuhan, China).

Techniques:

Journal: Scientific Reports

Article Title: GATA6 coordinates cross-talk between BMP10 and oxidative stress axis in pulmonary arterial hypertension

doi: 10.1038/s41598-023-33779-8

Figure Lengend Snippet: GATA6 loss induces SOD2 and other anti-oxidant enzymes deficiency in human and mouse PAECs. ( A ) An expression of the antioxidant enzymes measured by qPCR in human PAECs (HPAECs) transfected with siGATA6 or control scr siRNA. mRNA levels in scr siRNA transfected cells were set as one. Data are means ± SE, *p < 0.05 vs scr siRNA by Kruskal–Wallis test with post hoc Dunn’s test for multiple comparisons; n = 3–10 (see also Supplemental Table ). ( B ) Chromatin immune precipitation (ChiP) assay in HPAECs, n = 8 for SOD2 ; n = 5 for GPX1 . The experiments were repeated three times. Representative gel images and data quantification are shown. Values are means ± SE, **p < 0.01, ***p < 0.001 by Mann Whitney U test. ( C,D ) Control HPAECs transfected with siGATA6 or scr siRNA (siContr) were assayed by Western blot analysis to detect GATA6 and SOD2. Representative blots are shown. Values are means ± SE of the relative protein levels by densitometry, n = 5, **p < 0.01 by Mann Whitney U test. ( E,F ) Western blot analysis of whole lung tissue from Gata6 CKO and WT mice. Representative blots are shown. Values are means ± SE of the relative protein levels by densitometry, n = 4, *p < 0.05 by Mann Whitney U test. ( G ) SOD2, GPX, and Catalase activity were measured in Human HPAECs transfected with siGATA6 or scr siRNA, Values are means ± SE, n = 3, *p < 0.05 by Mann Whitney U test. ( H ) Mouse Sod2 (n = 5), Gpx (n = 10), and Cat (n = 9) activity were measured in mouse lung tissue in Gata6 CKO and WT mice. *p < 0.05, **p < 0.01 by Mann Whitney U test (Sod activity) and unpaired τ test (Gpx and Cat activities). ( I,J ) mRNA levels of the antioxidant enzymes measured by qPCR in whole lung tissues ( I ) and in PAEC from Gata6 CKO mice (J). Data are means ± SE. I: n = 12–13 for Sod , n = 10/group for Gpx1 , n = 9/group for Cat . J: n = 4 mice/group. *p < 0.05, **p < 0.01 by unpaired τ test (I) or Mann Whitney U test ( J ). The original blots are presented in Supplementary Fig. .

Article Snippet: Antibodies were as follows: Goat anti-human GATA6 (R&D; AF1700, 1:500), mouse anti-human GATA6 (Santa-Cruz, 517554, 1:500), Rabbit anti-human/mouse SOD2 (Cell signaling technology; 13141, 1:1000; 13145, 1:5000), Goat anti-human ALK1 (R&D; AF370, 1:500), Rabbit anti-mouse ALK1 (ABGENT AP7807a, 1:1000), rabbit anti-human/mouse ActR2B (LS bio; LS-B7781, 1:1000), mouse anti-human BMPR2 (Novus; NBP2-37624 Clone 3F6; 1:1000), rabbit anti-mouse BMPR2 (Proteintech; 19087–1-AP; 1:1000), mouse anti-mouse/human/rat BMPR2 (Abcam, catalog #130206 1:500), rabbit anti-human/mouse Endoglin (Proteintech; 10862–1-AP; 1:1000), rabbit anti-mouse BiP (Abcam; ab53068; 1:1000), rabbit anti-mouse CHOP (Novus; NBP2-13172; 1:1000), mouse anti-beta-actin (Sigma; A1978; 1:2000), mouse anti-human PRPF4 (Abcam, 69878, 1:1000), rabbit anti-human MYEOV (Invitrogen, PA5-48938 1:1000), rabbit anti-human STING (Invitrogen, PA5-26751, 1:1000), rabbit anti-human/mouse/rat EAF1 (Invitrogen, PA5-100493, 1:1000), rabbit anti-human/mouse/rat Histone H3 (Cell Signaling, 9715 1:1000), rabbit anti-human/mouse/rat a-b Tubulin (Cell Signaling, 2148, 1:1000).

Techniques: Expressing, Transfection, Control, MANN-WHITNEY, Western Blot, Activity Assay

Journal: Scientific Reports

Article Title: GATA6 coordinates cross-talk between BMP10 and oxidative stress axis in pulmonary arterial hypertension

doi: 10.1038/s41598-023-33779-8

Figure Lengend Snippet: GATA6 is deficient in both PAEC and PASMC in human PAH lungs. ( A ) IHC analyses were performed to detect GATA6 (green), CD31 (red) and α-SMA (red) in small PAs from healthy controls (HC) and patients with PAH (SSc-PAH and IPAH) and analysis of nuclear GATA6 in CD31- and SMA-positive cells in small PAs was performed. Left: Images are representative from 3–4 subjects/control, SSc-PAH, and IPAH. Bar equals 50 μm. White arrowheads indicate GATA6-positive cells. Right: Data are in optical density units (OD); Data are means ± SE from 3–4 human subjects per control, 7–8 for PAH (SSc-PAH + IPAH) groups. *p < 0.05 by Mann Whitney U test. ( B,C ) Immunoblot analysis of early-passage PAEC and PASMC from healthy controls (HC) and subjects with IPAH to detect indicated proteins. Data are means ± SE from n = 5 subjects/group. **p < 0.01 by Mann Whitney U test. ( D ) Immunoblot analysis of nuclear fractions of PASMC from healthy control (HC) and IPAH subjects. Data are means ± SE from n = 4 subjects/group. *p < 0.05 by Mann Whitney U test. ( E ) Immunocytochemical analysis of PASMC from healthy control (HC) and IPAH subjects to detect GATA6 (red) and DAPI (blue). Bar equals 50 µm. Representative images from two subjects/group by Mann Whitney U test. ( F–J ) Human non-diseased PASMC were transfected with siRNA GATA6 or control scrambled siRNA ( − ). 48 h post-transfection, immunoblot (F, G) and cell count analysis (H) were performed. Data are means ± SE from n = 3 subjects/group. *p < 0.05 by Mann Whitney U test. ( I ) qPCR analysis of PASMC from healthy controls (HC) and patients with PAH to detect SOD2 expression. Data are means ± SE, n = 5 subjects/group. *p < 0.05 by Mann Whitney U test. ( K,L ) Immunoblot analysis of healthy control (HC) and IPAH PASMC to detect indicated proteins was performed on the same membrane using strip- re-probe approach. Correlation between GATA6 and SOD was examined (R2). Data are means ± SE, n = 5 subjects/group. **p < 0.01 by Mann Whitney U test ( K ), R2 = 0.81 by Spearman analysis with Holm–Sidak adjusted p values ( L ). The original blots are presented in Supplementary Fig. .

Article Snippet: Antibodies were as follows: Goat anti-human GATA6 (R&D; AF1700, 1:500), mouse anti-human GATA6 (Santa-Cruz, 517554, 1:500), Rabbit anti-human/mouse SOD2 (Cell signaling technology; 13141, 1:1000; 13145, 1:5000), Goat anti-human ALK1 (R&D; AF370, 1:500), Rabbit anti-mouse ALK1 (ABGENT AP7807a, 1:1000), rabbit anti-human/mouse ActR2B (LS bio; LS-B7781, 1:1000), mouse anti-human BMPR2 (Novus; NBP2-37624 Clone 3F6; 1:1000), rabbit anti-mouse BMPR2 (Proteintech; 19087–1-AP; 1:1000), mouse anti-mouse/human/rat BMPR2 (Abcam, catalog #130206 1:500), rabbit anti-human/mouse Endoglin (Proteintech; 10862–1-AP; 1:1000), rabbit anti-mouse BiP (Abcam; ab53068; 1:1000), rabbit anti-mouse CHOP (Novus; NBP2-13172; 1:1000), mouse anti-beta-actin (Sigma; A1978; 1:2000), mouse anti-human PRPF4 (Abcam, 69878, 1:1000), rabbit anti-human MYEOV (Invitrogen, PA5-48938 1:1000), rabbit anti-human STING (Invitrogen, PA5-26751, 1:1000), rabbit anti-human/mouse/rat EAF1 (Invitrogen, PA5-100493, 1:1000), rabbit anti-human/mouse/rat Histone H3 (Cell Signaling, 9715 1:1000), rabbit anti-human/mouse/rat a-b Tubulin (Cell Signaling, 2148, 1:1000).

Techniques: Control, MANN-WHITNEY, Western Blot, Transfection, Cell Counting, Expressing, Membrane, Stripping Membranes

Journal: Scientific Reports

Article Title: GATA6 coordinates cross-talk between BMP10 and oxidative stress axis in pulmonary arterial hypertension

doi: 10.1038/s41598-023-33779-8

Figure Lengend Snippet: Treatment with DMF restores expression of the BMP receptors, reverses oxidative stress and pulmonary hypertension in Gata6 CKO mice. (DMF or vehicle were administered daily via i.p. injection for 3 weeks. ( A ) qPCR analysis of whole lung tissue from WT and Gata6 CKO mice treated with DMF or vehicle to detect expression of indicated BMP receptors. Data are means ± SE, n = 6–12, *p < 0.05. **p < 0.01 by Kruskal–Wallis test followed by Dunn’s multiple comparisons test ( BmpR2, ActRIIB, Alk1 ) and one-way ANOVA followed by post hoc Tukey’s multiple comparison ( Endoglin ). ( B ) qPCR analysis of whole lung tissue from WT and Gata6 CKO mice treated with DMF or vehicle to detect expression of the antioxidant enzymes and eNOS . Data are means ± SE, n = 6–17, *p < 0.05., **p < 0.01, ***p < 0.001 by one-way ANOVA followed by post hoc Tukey’s multiple comparisons test ( SOD2, GPX1, CAT ) and Kruskal–Wallis test with post hoc Dunn’s multiple comparisons test ( eNOS ). ( C,D ) mRNA levels of indicated BMP receptors and antioxidant enzymes measured by qPCR in PAEC from WT and Gata6 CKO mice treated with DMF or vehicle. Data are means ± SE, n = 3–6 mice/group, *p < 0.05, **p < 0.01 by Kruskal–Wallis test with post hoc Dunn’s multiple comparisons test. ( E–G ) RVSP, pulmonary acceleration time as a fraction of ejection time (PAT/ET) and Fulton index (RV/[LV + S]) were evaluated in WT and Gata6 CKO mice in the presence or absence of DMF. Data are means ± SE. n = 5–11 mice/group. *p < 0.05, **p < 0.01. ***p < 0.001 by Kruskal–Wallis test with post hoc Dunn’s multiple comparisons test (RVSP) and one-way ANOVA followed by post hoc Tukey’s multiple comparisons test (PAT/ET and RV/(LV + S).

Article Snippet: Antibodies were as follows: Goat anti-human GATA6 (R&D; AF1700, 1:500), mouse anti-human GATA6 (Santa-Cruz, 517554, 1:500), Rabbit anti-human/mouse SOD2 (Cell signaling technology; 13141, 1:1000; 13145, 1:5000), Goat anti-human ALK1 (R&D; AF370, 1:500), Rabbit anti-mouse ALK1 (ABGENT AP7807a, 1:1000), rabbit anti-human/mouse ActR2B (LS bio; LS-B7781, 1:1000), mouse anti-human BMPR2 (Novus; NBP2-37624 Clone 3F6; 1:1000), rabbit anti-mouse BMPR2 (Proteintech; 19087–1-AP; 1:1000), mouse anti-mouse/human/rat BMPR2 (Abcam, catalog #130206 1:500), rabbit anti-human/mouse Endoglin (Proteintech; 10862–1-AP; 1:1000), rabbit anti-mouse BiP (Abcam; ab53068; 1:1000), rabbit anti-mouse CHOP (Novus; NBP2-13172; 1:1000), mouse anti-beta-actin (Sigma; A1978; 1:2000), mouse anti-human PRPF4 (Abcam, 69878, 1:1000), rabbit anti-human MYEOV (Invitrogen, PA5-48938 1:1000), rabbit anti-human STING (Invitrogen, PA5-26751, 1:1000), rabbit anti-human/mouse/rat EAF1 (Invitrogen, PA5-100493, 1:1000), rabbit anti-human/mouse/rat Histone H3 (Cell Signaling, 9715 1:1000), rabbit anti-human/mouse/rat a-b Tubulin (Cell Signaling, 2148, 1:1000).

Techniques: Expressing, Injection, Comparison