Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Traditional Chinese Medicine Tang-Luo-Ning Ameliorates Sciatic Nerve Injuries in Streptozotocin-Induced Diabetic Rats

doi: 10.1155/2013/989670

Figure Lengend Snippet: Effect of TLN and α -LA on Nrg1/ErbB2 downstream signal pathway in sciatic nerve of streptozotocin-induced rats. A representative western blot ((a), (c), (e), and (g)) and the grey value analyses of protein expression ((b), (d), (f), and (h)) in sciatic nerves from different experimental groups. ((a), (b)) Ratio of P-ErbB2 to T-ErbB2 (P/T); ((c), (d)) ratio of P-Erk to T-Erk (P/T); ((e), (f)) ratio of P-Bad (Ser112) to T-Bad (P/T); ((g), (h)) ratio of P-Akt to T-Akt (P/T). Statistical analysis was performed using the ANOVA procedure t test (LSD). Data are expressed as mean ± SD. # P < 0.05 versus CTL group, * P < 0.05 versus STZ group. n = 4 per group. The protein expression of ratio of ErbB2 Erk, Bad (P/T) decreased compared with CTL group ( P < 0.05), while Tang-Luo-Ning and alpha-lipoic acid intervention could increase the ratio above compared with STZ group ( P < 0.05). There were no significant differences of ratio of Akt (P/T) between the four groups. Four groups: CTL group: nonstreptozotocin-induced group, STZ group: streptozotocin-induced diabetic group, TLN group: Tang-Luo-Ning group, and α -LA group: alpha-lipoic acid group. ANOVA: analysis of variance; LSD- t test: least significant difference t test.

Article Snippet: Rabbit polyclonal anti-ErbB2 antibody (dilution: 1 : 4000, Novus, Biologicals, Inc.; Littleton, CO, USA), Rabbit polyclonal anti-Phospho-ErbB2 (Tyr1248) antibody (dilution: 1 : 3000, Novus, Biologicals, Inc.; Littleton, CO, USA), after washing 3 × 10 min with Tris-buffered saline, horseradish peroxidase conjugated secondary antibody (dilution 1 : 1000: P-Bad ser112/ser136, 1 : 2000: other antibodies) was added and incubated for 1 hr with shaking.

Techniques: Western Blot, Expressing

Journal: Cancer & Metabolism

Article Title: Treatment of ErbB2 breast cancer by mitochondrial targeting

doi: 10.1186/s40170-020-00223-8

Figure Lengend Snippet: Suppression of ErbB2 breast cancer by MEDICA in vivo. a – c FVB-MMTV-ErbB2 transgenic mice were treated by MEDICA in feed as described in the “Methods” section. * Significant as compared to nontreated. a Breast tumor number (left), weight (middle), and volume (right). Mean ( n = 5–6/group). b Tumor ErbB1, ErbB2, phospho-ErbB2(Tyr1248), and ErbB3. Representative blots. c Lung rat ErbB2/neu (V664G) transcript. Mean ( n = 9–11/group). d NOD/SCID mice xenografted with RH2111 ErbB2 cells were treated with MEDICA in feed as described in the “Methods” section. Tumor volume (left) and tumor weight (right). Mean ± SD ( n = 4–5/group). Significant as compared to nontreated ( P < 0.05)

Article Snippet: Anti-ErbB2, anti-ErbB3, anti-EGFR, anti-caveolin-1, anti-phospho-ERK(Tyr204), anti-ERK, anti-phospho-Stat3(Tyr705), and anti-NDRG1 antibodies were from Santa Cruz Biotechnology INC. Anti-phospho-EGFR(Tyr1068) and anti-phospho-ErbB2(Tyr1248) antibodies were from Abcam.

Techniques: In Vivo, Transgenic Assay

Journal: Cancer & Metabolism

Article Title: Treatment of ErbB2 breast cancer by mitochondrial targeting

doi: 10.1186/s40170-020-00223-8

Figure Lengend Snippet: Growth inhibition of ErbB2 breast cancer cells by MEDICA. a – d Human ErbB2 AU565and BT474 cells, mouse ErbB2 RH2111 cells, and non-tumorigenic MCF10 cells were treated with MEDICA as indicated. a Cell growth. Mean ± SD. b Anchorage-dependent colonies. c Anchorage-independent colonies. d BT474 spheroids were allowed to form for 24 h followed by treatment for 8 days with MEDICA. Medium was refreshed every 3 days. Spheroid viability assayed by acid phosphatase was inhibited by 90% at 175 μM MEDICA. e Growth (72 h) inhibition of trastuzumab-resistant ErbB2 BT474 cells by MEDICA (left). Resistance was confirmed by growth with increasing trastuzumab concentrations as indicated (right). Mean ± SD. *Significant as compared to control ( P < 0.05). f Cell cycle (24 h) suppression of AU565 cells by 200uM MEDICA. Mean ± SD. *Significant as compared to control ( P < 0.05). Inset—representative micrographs

Article Snippet: Anti-ErbB2, anti-ErbB3, anti-EGFR, anti-caveolin-1, anti-phospho-ERK(Tyr204), anti-ERK, anti-phospho-Stat3(Tyr705), and anti-NDRG1 antibodies were from Santa Cruz Biotechnology INC. Anti-phospho-EGFR(Tyr1068) and anti-phospho-ErbB2(Tyr1248) antibodies were from Abcam.

Techniques: Inhibition

Journal: Cancer & Metabolism

Article Title: Treatment of ErbB2 breast cancer by mitochondrial targeting

doi: 10.1186/s40170-020-00223-8

Figure Lengend Snippet: Lipid raft disruption and loss of ErbB members by MEDICA. a AU565 cells were treated for 24 h with MEDICA as indicated. Plasma membrane ErbB2 and EGFR were determined by immunofluorescence confocal microscopy (red). Scale bar 20 μm. b AU565 cells were treated for 24 h with 200uM MEDICA. Plasma membrane ErbB members were tagged with Sulfo-NHS-Biotin and isolated by Streptavidin-Agarose beads. Plasma membrane (pm) and total ErbB members were analyzed by Western blot. c , d Lipid raft disruption by MEDICA. AU565 and BT474 cells were treated for 24 h and 40 h with 200uM MEDICA, respectively. Caveolin-1 was determined by FACS and by immunofluorescence confocal microscopy. GM1 was determined by immunofluorescence confocal microscopy. *Significant as compared to control ( P < 0.05). e AU565 cells were treated for 24 h with 200 μM MEDICA. Cell fractions prepared by density gradient centrifugation were subjected to GM1 (dot blot, upper frame) and ErbB2 (Western blot, lower frame) determination. Representative blots

Article Snippet: Anti-ErbB2, anti-ErbB3, anti-EGFR, anti-caveolin-1, anti-phospho-ERK(Tyr204), anti-ERK, anti-phospho-Stat3(Tyr705), and anti-NDRG1 antibodies were from Santa Cruz Biotechnology INC. Anti-phospho-EGFR(Tyr1068) and anti-phospho-ErbB2(Tyr1248) antibodies were from Abcam.

Techniques: Immunofluorescence, Confocal Microscopy, Isolation, Western Blot, Gradient Centrifugation, Dot Blot

Journal: Diabetes

Article Title: Caveolin-1 and Altered Neuregulin Signaling Contribute to the Pathophysiological Progression of Diabetic Peripheral Neuropathy

doi: 10.2337/db09-0594

Figure Lengend Snippet: Diabetes increases pErb B2 in sciatic nerve. A , C , and E : Upper panels show representative immunoblot analysis demonstrating induction of pErb B2 in diabetic mice after 2, 6, and 12 weeks of diabetes, respectively. B , D , and E : Immmunoblot quantitation of Erb B2 and pErb B2 after 2, 6, and 12 weeks of diabetes, respectively (nondiabetic wild-type and Cav1 knockout [ n = 3–4]; diabetic wild-type and Cav1 knockout [ n = 4–6]). Levels of Erb B2 and pErb B2 were normalized to β-actin and expressed as a percent of the genotype control. * P < 0.05 compared with genotype control. ∧ P < 0.05 compared with diabetic wild-type mice. G : Immunofluorescence analysis of pErb B2 induction in cross section of sciatic nerve of 6-week-diabetic wild-type mouse. NF-H, neurofilament heavy chain. KO, knockout; WT, wild type. (A high-quality color digital representation of this figure is available in the online issue.)

Article Snippet: The antibodies used and their sources were: Cav1 2234 (Transduction Labs, Lexington, KY); Erb B2 (Millipore, Billerica, MA); phospho-Tyr 1248 Erb B2 (pErb B2), β-actin, neurofilament H, and horseradish peroxidase–conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA); and AlexaFluor 488 rabbit anti-mouse and AlexaFluor 568 goat anti-rabbit antibodies (Molecular Probes, Eugene, OR).

Techniques: Western Blot, Quantitation Assay, Knock-Out, Control, Immunofluorescence

Journal: Diabetes

Article Title: Caveolin-1 and Altered Neuregulin Signaling Contribute to the Pathophysiological Progression of Diabetic Peripheral Neuropathy

doi: 10.2337/db09-0594

Figure Lengend Snippet: Verification of the tissue specificity of the rtTA transgene and generation of P 0 -rtTA × caErb B2 bitransgenics. A : mRNA was isolated from various tissues obtained from a progeny of one founder line, and cDNA was prepared. The rtTA transcript was amplified by PCR using 2 and 4 μg of total cDNA. The β-actin transcript was amplified by PCR using 2 μg of total cDNA. The rtTA transcript was enriched in samples from sciatic nerve despite a low level of β-actin. Br, brain; H, heart; K, kidney; Li, liver; Lu, lung; M, muscle; SN, sciatic nerve; Sp, spleen. B : Schematic of PCR strategy for identifying P 0 -rtTa and TRE-caErb B2 transgenes. C : Identification of several bitransgenic (biTg) and one nonbiTg progeny from cross of P 0 -rtTa and TRE-caErb B2 parents. Bottom panel shows the presence of transgenes in the parents but their absence in a wild-type (WT) mouse. Amplicons identify the TRE-caErb B2 ( lane a ) and P 0 -rtTA ( lane b ) transgenes. Lane c is positive control for the presence of the endogenous P 0 promoter.

Article Snippet: The antibodies used and their sources were: Cav1 2234 (Transduction Labs, Lexington, KY); Erb B2 (Millipore, Billerica, MA); phospho-Tyr 1248 Erb B2 (pErb B2), β-actin, neurofilament H, and horseradish peroxidase–conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA); and AlexaFluor 488 rabbit anti-mouse and AlexaFluor 568 goat anti-rabbit antibodies (Molecular Probes, Eugene, OR).

Techniques: Isolation, Amplification, Positive Control

Journal: Diabetes

Article Title: Caveolin-1 and Altered Neuregulin Signaling Contribute to the Pathophysiological Progression of Diabetic Peripheral Neuropathy

doi: 10.2337/db09-0594

Figure Lengend Snippet: Induction of caErb B2 in sciatic nerve of bitransgenic mice. A : Bitransgenic mice were placed on standard rat chow (−) or the DOX diet (+) for 9 weeks. Sciatic nerves were harvested, and immunoblot analysis was performed for pErb B2, Erb B2, and β-actin. B : Bitransgenic mice were placed on standard rat chow (−) or the DOX diet (+) for 9 weeks, and the DOX diet was replaced (±) with standard rat chow for 3 weeks. Sciatic nerves were harvested, and immunoblot analysis was performed for pErb B2 and Erb B2.

Article Snippet: The antibodies used and their sources were: Cav1 2234 (Transduction Labs, Lexington, KY); Erb B2 (Millipore, Billerica, MA); phospho-Tyr 1248 Erb B2 (pErb B2), β-actin, neurofilament H, and horseradish peroxidase–conjugated secondary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA); and AlexaFluor 488 rabbit anti-mouse and AlexaFluor 568 goat anti-rabbit antibodies (Molecular Probes, Eugene, OR).

Techniques: Western Blot