Journal: Biology

Article Title: MYOC Promotes the Differentiation of C2C12 Cells by Regulation of the TGF-β Signaling Pathways via CAV1

doi: 10.3390/biology10070686

Figure Lengend Snippet: Effects of CAV1 on C2C12 cell differentiation. ( A ). Change in CAV1, MYH2, and MYOG expression levels at different stages of C2C12 cell differentiation. ( B ). Grayscale scan of CAV1 in Figure A. ( C ). Grayscale scan of MYH2 in Figure A. ( D ). Grayscale scan of MYOG in Figure A. ( E ). Immunofluorescence was used to measure the changes in CAV1 expression levels and the morphology of myotubes at various stages of C2C12 differentiation. CAV1 was labeled green using FITC and the nucleus blue using DAPI. ( F ). Laser confocal microscopy demonstrating the location of CAV1 expression, stained red using RBFITC, and nuclei blue using DAPI. ( G ). Changes in CAV1, MYH2, and MYOG expression levels after siRNA transfection for 48 h and 72 h. ( H ). Grayscale scan of CAV1 in Figure G. ( I ). Grayscale scan of MYH2 in Figure G. ( J ). Grayscale scan of MYOG in Figure G. ( K ). Changes in the fusion rate of C2C12 cells in mice after CAV1 inhibition by immunofluorescence detection. ( L ). Myotubule fusion rate in Figure K. ( M ). Change in CAV1, P-Smad2, and P-Smad4 expression after siRNA fragment S1 transfection of CAV1. ( N ). Grayscale scan of CAV1 in Figure M. ( O ). Grayscale scan of P-Smad2 in Figure M. ( P ). Grayscale scan of P-Smad4 in Figure M. (** p values < 0.01, * p values < 0.05) ( n = 3).

Article Snippet: Antibodies used for Western blotting were: anti-MYOC (1:1000, Sangon, D154098, Shanghai, China), anti-CAV1 (1:1000, Sangon, D261423), anti-MYH2 (1:1000, Santa Cruz Biotechnology, SC-53092, Dallas, TX, USA), anti-MYOG (1:500, Santa, SC-12732), anti-GAPDH (1:2000, Proteintech, 10494-1-AP, Rosemont, IL, USA), anti-Smad2 (1:1000, bs-0718R, Bioss, Beijing, China), anti-P-Smad2 (1:500, bs-7464R, Bioss), anti-Smad4 (1:1000, D120124, Sangon), and anti-P-Smad4 (1:500, Affinity, Ab-AF8316, Jiangsu, China).

Techniques: Cell Differentiation, Expressing, Immunofluorescence, Labeling, Confocal Microscopy, Staining, Transfection, Inhibition

Journal: Biology

Article Title: MYOC Promotes the Differentiation of C2C12 Cells by Regulation of the TGF-β Signaling Pathways via CAV1

doi: 10.3390/biology10070686

Figure Lengend Snippet: MYOC exerts an effect on the differentiation of C2C12 cells by regulation of CAV1 that affects the TGF-β pathway. ( A ). Change in MYOC, P-Smad2, and P-SMad4 expression after transfection of siRNA-sequence T3 for MYOC silencing. ( B ). Grayscale scan of MYOC in Figure A. ( C ). Grayscale scan of P-Smad2 in Figure A. ( D ). Grayscale scan of P-Smad4 in Figure A. ( E ). Western blotting of all indices in the co-transfection experiment control group and 3 experimental groups. ( F ). Grayscale scan of MYOC in Figure E. ( G ). Grayscale scan of CAV1 in Figure E. ( H ). Grayscale scan of MYH2 in Figure E. ( I ). Grayscale scan of MYOG in Figure E. ( J ). Grayscale scan of P-Smad2 in Figure E. ( K ). Grayscale scanning of P-Smad4 in Figure E. ( L ). Change in rate of fusion of myotubes after co-transfection, as measured by laser confocal microscopy. ( M ). Myotubule fusion rate in Figure L. (** p values < 0.01, * p values < 0.05) ( n = 3).

Article Snippet: Antibodies used for Western blotting were: anti-MYOC (1:1000, Sangon, D154098, Shanghai, China), anti-CAV1 (1:1000, Sangon, D261423), anti-MYH2 (1:1000, Santa Cruz Biotechnology, SC-53092, Dallas, TX, USA), anti-MYOG (1:500, Santa, SC-12732), anti-GAPDH (1:2000, Proteintech, 10494-1-AP, Rosemont, IL, USA), anti-Smad2 (1:1000, bs-0718R, Bioss, Beijing, China), anti-P-Smad2 (1:500, bs-7464R, Bioss), anti-Smad4 (1:1000, D120124, Sangon), and anti-P-Smad4 (1:500, Affinity, Ab-AF8316, Jiangsu, China).

Techniques: Expressing, Transfection, Sequencing, Western Blot, Cotransfection, Confocal Microscopy

Journal: bioRxiv

Article Title: Enrichment of breast cancer stem cells following cytotoxic chemotherapy is mediated by hypoxia-inducible factors

doi: 10.1101/2022.06.27.497729

Figure Lengend Snippet: NTC and DKD subclones were treated with either vehicle or Pac for four days and IB assays were performed using antibodies specific for phosphorylated (p-) and total (t-) SMAD2 ( A ) or STAT3 ( B ).

Article Snippet: Blots were probed with antibodies against HIF-1α, HIF-2α, IL-6, IL-8, MDR1, phospho-SMAD2, phospho-STAT3, SMAD2, and STAT3 (Novus Biologicals), followed by HRP-conjugated anti-rabbit (Roche) or anti-mouse (Santa Cruz) secondary antibodies and signal was detected using ECL Plus (GE Healthcare).

Techniques:

Journal: Biochemical Journal

Article Title: Connective tissue growth factor antagonizes transforming growth factor-β1/Smad signalling in renal mesangial cells

doi: 10.1042/bj20110910

Figure Lengend Snippet: Figure 1 CTGF inhibits TGF-β1-induced Smad2/3 phosphorylation, 3TP-lux promoter activity and TGF-β1 receptor binding

Article Snippet: Antibodies against phospho-p44/42 MAPK (mitogenactivated protein kinase; Thr202/Tyr204), total p44/42 MAPK, phospho-Smad2 (Ser465/Ser467), total Smad2, phospho-Smad3 (Ser423/Ser425), total Smad3, phospho-Smad2 (Ser245/Ser250/Ser255), phospho-Smad1 (Ser463/Ser465)/Smad5 (Ser463/Ser465)/Smad8 (Ser426/Ser428), total Smad5 and TβRIII (#2519) were from Cell Signaling Technology.

Techniques: Phospho-proteomics, Activity Assay, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: The Ethyl Acetate Extract of Phyllanthus emblica L. Alleviates Diabetic Nephropathy in a Murine Model of Diabetes

doi: 10.3390/ijms25126686

Figure Lengend Snippet: The kidney target gene expression levels in db/db mice following treatment with ethyl acetate extract of P. emblica L. (EPE) or aminoguanidine (AG, 20 mg/kg body weight) by Western blotting analysis on p-PKCα/t-PKCα, VEGF, nephrin, TGFβ1, collagen IV, fibronectin, Smad4, p-Smad2/t-Smad2, and p-Smad3/t-Smad3. ( A , C , E , G ) Representative image; ( B , D , F , H ) quantification of p-PKCα/t-PKCα, VEGF TGFβ1, collagen IV, Smad4, p-Smad2/t-Smad2, p-Smad3/t-Smad3, p-NLRP3/t-NLRP3, ICAM, and Capalase-1 to β-actin. Protein was separated by 12% SDS-PAG. ### p < 0.001 as compared to the db/m group; * p < 0.05, ** p < 0.01, *** p < 0.001 compared to the db/db plus vehicle (distilled water) (db/db) group. All values are means ± SE ( n = 8 per group). EPE, ethyl acetate extract of P. emblica L. EPE: EPE1: 100, EPE2: 200, EPE3: 400 mg/kg body weight; AG: aminoguanidine (20 mg/kg body weight).

Article Snippet: Antibodies against Smad-2 (no. GTX111075), Smad-3 (no. GTX34208), Smad-4 (no. GTX112980), phospho-Smad2 (Ser250) [GT1291] (no. GTX03203), and phospho-Smad3 (Ser423/425) (no. GTX129841) were obtained from GeneTex International Corporation (East Dist., Hsinchu City, Taiwan).

Techniques: Targeted Gene Expression, Western Blot