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Effect of CKD on early induced preferential atrophy of oxidative muscle. Evaluation of skeletal muscle atrophy in CKD mice. (A) The experimental scheme. A combination of 2/3 Nx+UUO was performed using a two-step surgery. The mice were sacrificed, and an evaluation was performed 4 weeks after the final surgery. (B) Renal function, (C) body weight, (D) grip strength, and (E) muscle weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed 4 weeks after the final surgery. (F) Plasma <t>TGF-</t> β levels were determined by ELISA. (G and H) Distribution of muscle fiber CSA for (G) oxidative and (H) glycolytic muscles was quantitatively analyzed using a BZ-X analyzer. Data are expressed as means±SEM ( n =6). * P < 0.05, ** P < 0.01 compared with the sham group. 2/3 Nx+UUO, 2/3 nephrectomy and unilateral ureteral obstruction; CSA, cross-sectional area.

Tgf β R2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 248 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tgf+%CE%B2+r2+antibodies/TGF%CE%B2+RII+Antibody/pmc12483047-112-29-32
Average 94 stars, based on 248 article reviews

tgf β r2 - by Bioz Stars, 2026-10

94/100 stars

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1) Product Images from "CKD-Induced Oxidative Twitch Muscle Atrophy Is Mediated by TGF- β"

Article Title: CKD-Induced Oxidative Twitch Muscle Atrophy Is Mediated by TGF- β

Journal: Kidney360

doi: 10.34067/KID.0000000852

Figure Legend Snippet: Effect of CKD on early induced preferential atrophy of oxidative muscle. Evaluation of skeletal muscle atrophy in CKD mice. (A) The experimental scheme. A combination of 2/3 Nx+UUO was performed using a two-step surgery. The mice were sacrificed, and an evaluation was performed 4 weeks after the final surgery. (B) Renal function, (C) body weight, (D) grip strength, and (E) muscle weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed 4 weeks after the final surgery. (F) Plasma TGF- β levels were determined by ELISA. (G and H) Distribution of muscle fiber CSA for (G) oxidative and (H) glycolytic muscles was quantitatively analyzed using a BZ-X analyzer. Data are expressed as means±SEM ( n =6). * P < 0.05, ** P < 0.01 compared with the sham group. 2/3 Nx+UUO, 2/3 nephrectomy and unilateral ureteral obstruction; CSA, cross-sectional area.

Techniques Used: Muscles, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Effect of TGF- β for preferentially induced oxidative muscle atrophy. Evaluation of skeletal muscle atrophy in TGF- β –treated mice. (A) The experimental scheme. TGF- β was administered via intraperitoneal injection at a dose of 50 ng/d. Mice in the normal group were intraperitoneally injected with equivalent amounts of saline daily. Both groups were treated for 4 weeks. (B) Renal function, (C) body weight, (D) grip strength, and (E) weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed at 4 weeks. (F) Representative images of coimmunostaining of skeletal muscle phenotype (top: MyHC 1, blue; MyHC 2a, green; MyHC 2b, red; bottom: MyHC 2a, green; MyHC 2x, red) markers in the soleus and deep tibialis anterior muscle sections from sham and TGF- β– treated mice. Scale bars: 100 μ m. Images were captured at 200× magnification. (G) Protein levels of PGC-1 α , p-SMAD2/3, and SMAD2/3 as determined by Western blot. Data are expressed as means±SEM ( n =6). * P < 0.05, ** P < 0.01 compared with the sham group. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MyHC, myosin heavy chain; PGC-1 α , peroxisome proliferator-activated receptor-γ coactivator-1 α .

Figure Legend Snippet: Effect of TGF- β for preferentially induced oxidative muscle atrophy. Evaluation of skeletal muscle atrophy in TGF- β –treated mice. (A) The experimental scheme. TGF- β was administered via intraperitoneal injection at a dose of 50 ng/d. Mice in the normal group were intraperitoneally injected with equivalent amounts of saline daily. Both groups were treated for 4 weeks. (B) Renal function, (C) body weight, (D) grip strength, and (E) weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed at 4 weeks. (F) Representative images of coimmunostaining of skeletal muscle phenotype (top: MyHC 1, blue; MyHC 2a, green; MyHC 2b, red; bottom: MyHC 2a, green; MyHC 2x, red) markers in the soleus and deep tibialis anterior muscle sections from sham and TGF- β– treated mice. Scale bars: 100 μ m. Images were captured at 200× magnification. (G) Protein levels of PGC-1 α , p-SMAD2/3, and SMAD2/3 as determined by Western blot. Data are expressed as means±SEM ( n =6). * P < 0.05, ** P < 0.01 compared with the sham group. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MyHC, myosin heavy chain; PGC-1 α , peroxisome proliferator-activated receptor-γ coactivator-1 α .

Techniques Used: Injection, Saline, Muscles, Western Blot

Effect of TGF- β inh. on CKD-induced oxidative muscle atrophy and exercise capacity in CKD mice. Evaluation of skeletal muscle atrophy in TGF- β inh.–treated CKD mice. (A) The experimental scheme. TGF- β inh. was orally administered at a dose of 3 mg/kg per day. Mice in the sham and CKD groups were orally administered equivalent amounts of 0.5% carboxymethylcellulose sodium daily. All groups were treated for 4 weeks. (B) Renal function, (C) body weight, (D) grip strength, and (E) weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed at 4 weeks. (F) Representative images of coimmunostaining of skeletal muscle phenotype (top: MyHC 1, blue; MyHC 2a, green; MyHC 2b, red; bottom: MyHC 2a, green; MyHC 2x, red) markers in the soleus and deep tibialis anterior sections from sham, CKD, and TGF- β inh.–treated CKD mice. Scale bars: 100 μ m. Images were captured at 200× magnification. (G) Protein levels of PGC-1 α , p-SMAD2/3, and SMAD2/3 as determined by Western blot. (H) Work and (I) running distances were measured using a treadmill running test. (J) Representative images of coimmunostaining of MyHC isoforms and TGFBR2 (top: MyHC 1, blue; MyHC 2a, green; TGFBR2, red; middle: TGFBR2, green; MyHC 2x, red; bottom: TGFBR2, green; MyHC 2b, red) markers in the soleus and deep tibialis anterior sections from sham and CKD mice. Scale bars: 50 μ m. The arrows (▲) indicate the distribution of TGFBR2. Images were captured at 400× magnification. Data are expressed as means±SEM ( n =5). * P < 0.05, ** P < 0.01 compared with the sham group. † P < 0.05 compared with the CKD group. TGF- β inh., TGF- β inhibitor; TGFBR2, TGF- β type 2 receptor.

Figure Legend Snippet: Effect of TGF- β inh. on CKD-induced oxidative muscle atrophy and exercise capacity in CKD mice. Evaluation of skeletal muscle atrophy in TGF- β inh.–treated CKD mice. (A) The experimental scheme. TGF- β inh. was orally administered at a dose of 3 mg/kg per day. Mice in the sham and CKD groups were orally administered equivalent amounts of 0.5% carboxymethylcellulose sodium daily. All groups were treated for 4 weeks. (B) Renal function, (C) body weight, (D) grip strength, and (E) weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed at 4 weeks. (F) Representative images of coimmunostaining of skeletal muscle phenotype (top: MyHC 1, blue; MyHC 2a, green; MyHC 2b, red; bottom: MyHC 2a, green; MyHC 2x, red) markers in the soleus and deep tibialis anterior sections from sham, CKD, and TGF- β inh.–treated CKD mice. Scale bars: 100 μ m. Images were captured at 200× magnification. (G) Protein levels of PGC-1 α , p-SMAD2/3, and SMAD2/3 as determined by Western blot. (H) Work and (I) running distances were measured using a treadmill running test. (J) Representative images of coimmunostaining of MyHC isoforms and TGFBR2 (top: MyHC 1, blue; MyHC 2a, green; TGFBR2, red; middle: TGFBR2, green; MyHC 2x, red; bottom: TGFBR2, green; MyHC 2b, red) markers in the soleus and deep tibialis anterior sections from sham and CKD mice. Scale bars: 50 μ m. The arrows (▲) indicate the distribution of TGFBR2. Images were captured at 400× magnification. Data are expressed as means±SEM ( n =5). * P < 0.05, ** P < 0.01 compared with the sham group. † P < 0.05 compared with the CKD group. TGF- β inh., TGF- β inhibitor; TGFBR2, TGF- β type 2 receptor.

Techniques Used: Muscles, Western Blot

$Determination of the major factor for MyHC 2a-selective atrophy using C2C12 myotubes. (A) C2C12 myoblasts were differentiated into myotubes for 5 days and then treated with sham or CKD mice serum for 3 days. Protein levels of skeletal muscle phenotype (MyHC 1, 2a, 2b, and 2x), as determined by Western blot. * P < 0.05, ** P < 0.01 compared with the sham. (B) Sham and CKD mice serum components were separated using a 3 kDa centrifugal ultrafiltration filter. C2C12 myoblasts were differentiated into myotubes for 5 days and then treated with fractionated serum components of sham or CKD mice for 3 days. Protein levels of skeletal muscle phenotype as determined by Western blot. Student t test was performed for the upper and lower layer sham and CKD groups. * P < 0.05, ** P < 0.01 compared with the upper layer of sham mouse serum. (C) C2C12 myotubes were pretreated with TGF- β inh. (R-268712) for 30 minutes and then treated with sham or CKD mice serum for 3 days. Protein levels of skeletal muscle phenotype as determined by western blot. Student t test was performed for the sham and CKD groups of TGF- β inh.–treated or untreated serum. * P < 0.05 compared with the untreated sham mouse serum. Data are expressed as means±SEM ( n =3).$
Figure Legend Snippet: Determination of the major factor for MyHC 2a-selective atrophy using C2C12 myotubes. (A) C2C12 myoblasts were differentiated into myotubes for 5 days and then treated with sham or CKD mice serum for 3 days. Protein levels of skeletal muscle phenotype (MyHC 1, 2a, 2b, and 2x), as determined by Western blot. * P < 0.05, ** P < 0.01 compared with the sham. (B) Sham and CKD mice serum components were separated using a 3 kDa centrifugal ultrafiltration filter. C2C12 myoblasts were differentiated into myotubes for 5 days and then treated with fractionated serum components of sham or CKD mice for 3 days. Protein levels of skeletal muscle phenotype as determined by Western blot. Student t test was performed for the upper and lower layer sham and CKD groups. * P < 0.05, ** P < 0.01 compared with the upper layer of sham mouse serum. (C) C2C12 myotubes were pretreated with TGF- β inh. (R-268712) for 30 minutes and then treated with sham or CKD mice serum for 3 days. Protein levels of skeletal muscle phenotype as determined by western blot. Student t test was performed for the sham and CKD groups of TGF- β inh.–treated or untreated serum. * P < 0.05 compared with the untreated sham mouse serum. Data are expressed as means±SEM ( n =3).

Techniques Used: Western Blot

Incubation:

Article Title: Predisposition for Disrepair in the Aged Lung
Article Snippet: .. The blots were blocked and then incubated with a rat anti-TGF- β 1 antibody (1:500; Abcam, Cambridge, MA), a rabbit anti-Fn antibody (1:5000; Sigma-Aldrich), TGF- β R1 (1:200 dilution) and TGF- β R2 antibodies (1:500; Santa Cruz Biotechnology, Santa Cruz, CA), Smad2/3 (1:1000; Cell Signaling, Danvers, MA) and phosphorylated Smad3 antibodies (1:1000; Cell Signaling), a mouse anti-Fn-EDA antibody (1:200; Abcam) or β -actin antibody (1:500; Sigma-Aldrich, St. Louis, MO) at 4 °C overnight. .. The blots were washed and incubated with an appropriate horseradish peroxidase– conjugated secondary antibody (at 1:2000 dilution for all blots; Amersham Biosciences, Pittsburgh, PA), washed and visualized through enzyme-linked chemiluminescence using the SuperSignal West Pico kit (Pierce Biotechnology, Rockford, IL).

Image Search Results

Journal: Kidney360

Article Title: CKD-Induced Oxidative Twitch Muscle Atrophy Is Mediated by TGF- β

doi: 10.34067/KID.0000000852

Figure Lengend Snippet: Effect of CKD on early induced preferential atrophy of oxidative muscle. Evaluation of skeletal muscle atrophy in CKD mice. (A) The experimental scheme. A combination of 2/3 Nx+UUO was performed using a two-step surgery. The mice were sacrificed, and an evaluation was performed 4 weeks after the final surgery. (B) Renal function, (C) body weight, (D) grip strength, and (E) muscle weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed 4 weeks after the final surgery. (F) Plasma TGF- β levels were determined by ELISA. (G and H) Distribution of muscle fiber CSA for (G) oxidative and (H) glycolytic muscles was quantitatively analyzed using a BZ-X analyzer. Data are expressed as means±SEM ( n =6). * P < 0.05, ** P < 0.01 compared with the sham group. 2/3 Nx+UUO, 2/3 nephrectomy and unilateral ureteral obstruction; CSA, cross-sectional area.

Article Snippet: Subsequently, sections were incubated with primary antibodies against laminin (sigma, cat#: L9393), MyHC 1 (DSHB, BA-F8), MyHC 2a (DSHB, SC-71), MyHC 2b (DSHB, BF-F3), MyHC 2x (DSHB, 6H1), and TGF- β R2 (Santa Cruz Biotechnology, sc-17791) and then labeled with Flexible CoraLite Plus 488 (Proteintech, KFA021) and Flexible CoraLite Plus 555 (Proteintech, KFA022) for 1 hour ( Supplemental Figure 5 and Supplemental Table 1 ).

Techniques: Muscles, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Kidney360

Article Title: CKD-Induced Oxidative Twitch Muscle Atrophy Is Mediated by TGF- β

doi: 10.34067/KID.0000000852

Figure Lengend Snippet: Effect of TGF- β for preferentially induced oxidative muscle atrophy. Evaluation of skeletal muscle atrophy in TGF- β –treated mice. (A) The experimental scheme. TGF- β was administered via intraperitoneal injection at a dose of 50 ng/d. Mice in the normal group were intraperitoneally injected with equivalent amounts of saline daily. Both groups were treated for 4 weeks. (B) Renal function, (C) body weight, (D) grip strength, and (E) weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed at 4 weeks. (F) Representative images of coimmunostaining of skeletal muscle phenotype (top: MyHC 1, blue; MyHC 2a, green; MyHC 2b, red; bottom: MyHC 2a, green; MyHC 2x, red) markers in the soleus and deep tibialis anterior muscle sections from sham and TGF- β– treated mice. Scale bars: 100 μ m. Images were captured at 200× magnification. (G) Protein levels of PGC-1 α , p-SMAD2/3, and SMAD2/3 as determined by Western blot. Data are expressed as means±SEM ( n =6). * P < 0.05, ** P < 0.01 compared with the sham group. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; MyHC, myosin heavy chain; PGC-1 α , peroxisome proliferator-activated receptor-γ coactivator-1 α .

Techniques: Injection, Saline, Muscles, Western Blot

Journal: Kidney360

Article Title: CKD-Induced Oxidative Twitch Muscle Atrophy Is Mediated by TGF- β

doi: 10.34067/KID.0000000852

Figure Lengend Snippet: Effect of TGF- β inh. on CKD-induced oxidative muscle atrophy and exercise capacity in CKD mice. Evaluation of skeletal muscle atrophy in TGF- β inh.–treated CKD mice. (A) The experimental scheme. TGF- β inh. was orally administered at a dose of 3 mg/kg per day. Mice in the sham and CKD groups were orally administered equivalent amounts of 0.5% carboxymethylcellulose sodium daily. All groups were treated for 4 weeks. (B) Renal function, (C) body weight, (D) grip strength, and (E) weights of the soleus, extensor digitorum longus, tibialis anterior, and gastrocnemius muscles were assessed at 4 weeks. (F) Representative images of coimmunostaining of skeletal muscle phenotype (top: MyHC 1, blue; MyHC 2a, green; MyHC 2b, red; bottom: MyHC 2a, green; MyHC 2x, red) markers in the soleus and deep tibialis anterior sections from sham, CKD, and TGF- β inh.–treated CKD mice. Scale bars: 100 μ m. Images were captured at 200× magnification. (G) Protein levels of PGC-1 α , p-SMAD2/3, and SMAD2/3 as determined by Western blot. (H) Work and (I) running distances were measured using a treadmill running test. (J) Representative images of coimmunostaining of MyHC isoforms and TGFBR2 (top: MyHC 1, blue; MyHC 2a, green; TGFBR2, red; middle: TGFBR2, green; MyHC 2x, red; bottom: TGFBR2, green; MyHC 2b, red) markers in the soleus and deep tibialis anterior sections from sham and CKD mice. Scale bars: 50 μ m. The arrows (▲) indicate the distribution of TGFBR2. Images were captured at 400× magnification. Data are expressed as means±SEM ( n =5). * P < 0.05, ** P < 0.01 compared with the sham group. † P < 0.05 compared with the CKD group. TGF- β inh., TGF- β inhibitor; TGFBR2, TGF- β type 2 receptor.

Techniques: Muscles, Western Blot

Journal: Kidney360

Article Title: CKD-Induced Oxidative Twitch Muscle Atrophy Is Mediated by TGF- β

doi: 10.34067/KID.0000000852

Figure Lengend Snippet: Determination of the major factor for MyHC 2a-selective atrophy using C2C12 myotubes. (A) C2C12 myoblasts were differentiated into myotubes for 5 days and then treated with sham or CKD mice serum for 3 days. Protein levels of skeletal muscle phenotype (MyHC 1, 2a, 2b, and 2x), as determined by Western blot. * P < 0.05, ** P < 0.01 compared with the sham. (B) Sham and CKD mice serum components were separated using a 3 kDa centrifugal ultrafiltration filter. C2C12 myoblasts were differentiated into myotubes for 5 days and then treated with fractionated serum components of sham or CKD mice for 3 days. Protein levels of skeletal muscle phenotype as determined by Western blot. Student t test was performed for the upper and lower layer sham and CKD groups. * P < 0.05, ** P < 0.01 compared with the upper layer of sham mouse serum. (C) C2C12 myotubes were pretreated with TGF- β inh. (R-268712) for 30 minutes and then treated with sham or CKD mice serum for 3 days. Protein levels of skeletal muscle phenotype as determined by western blot. Student t test was performed for the sham and CKD groups of TGF- β inh.–treated or untreated serum. * P < 0.05 compared with the untreated sham mouse serum. Data are expressed as means±SEM ( n =3).

Techniques: Western Blot

Figure 4. HA and CD44 are involved in rhHAPLN1-induced TGF-β R2 upregulation.

Journal: Matrix biology : journal of the International Society for Matrix Biology

Article Title: Hyaluronan and proteoglycan link protein 1 - a novel signaling molecule for rejuvenating aged skin.

doi: 10.1016/j.matbio.2024.08.009

Figure Lengend Snippet: Figure 4. HA and CD44 are involved in rhHAPLN1-induced TGF-β R2 upregulation.

Article Snippet: The cells were then incubated with phycoerythrin-conjugated CD44 IgG (BioLegend, USA, #103008) and Alexa Fluor 488-conjugated TGF-β R2 IgG (Santa Cruz, #SC-17799 AF488) on ice for 1 h. After washing, the cells were analyzed using a flow cytometer (BD Biosciences, USA).

Techniques:

In a mouse lung fibrosis model constructed by bleomycin, fibrosis was significantly attenuated with the intervention of pirfenidone and nintedanib. (A) Levels of TGF-β1, SP-A, SP-D and KL-6 in the plasma of mice. The plasma levels of TGF-β1, KL-6, SP-A and SP-D were significantly increased in mice and were inhibited by pirfenidone and nintedanib (n=3, # , P<0.05; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with BLM group). (B) Representative photographs of pirfenidone and nintedanib on bleomycin-induced pulmonary pathological changes in mice (H&E staining and Masson staining of mouse lung tissue). On day 28, interstitial fibrosis was evident in the lungs of mice in the Bleomycin group compared with the Control group. However, lung histopathological changes were significantly reduced in the Pirfenidone group, Nintedanib group. (C) Immunohistochemistry of α-SMA and collagen I (brown areas) in bleomycin-induced pulmonary fibrosis in mice on day 14. Representative images of lung sections from each group are shown. TGF, transforming growth factor; H&E, hematoxylin-eosin; SMA, smooth muscle actin; MOD, mean optical density; BLM, bleomycin.

Journal: Journal of Thoracic Disease

Article Title: Pirfenidone and nintedanib attenuate pulmonary fibrosis in mice by inhibiting the expression of JAK2

doi: 10.21037/jtd-23-1057

Figure Lengend Snippet: In a mouse lung fibrosis model constructed by bleomycin, fibrosis was significantly attenuated with the intervention of pirfenidone and nintedanib. (A) Levels of TGF-β1, SP-A, SP-D and KL-6 in the plasma of mice. The plasma levels of TGF-β1, KL-6, SP-A and SP-D were significantly increased in mice and were inhibited by pirfenidone and nintedanib (n=3, # , P<0.05; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with BLM group). (B) Representative photographs of pirfenidone and nintedanib on bleomycin-induced pulmonary pathological changes in mice (H&E staining and Masson staining of mouse lung tissue). On day 28, interstitial fibrosis was evident in the lungs of mice in the Bleomycin group compared with the Control group. However, lung histopathological changes were significantly reduced in the Pirfenidone group, Nintedanib group. (C) Immunohistochemistry of α-SMA and collagen I (brown areas) in bleomycin-induced pulmonary fibrosis in mice on day 14. Representative images of lung sections from each group are shown. TGF, transforming growth factor; H&E, hematoxylin-eosin; SMA, smooth muscle actin; MOD, mean optical density; BLM, bleomycin.

Article Snippet: Anti-TGF-β-R2 antibodies were purchased from Proteintech.

Techniques: Construct, Clinical Proteomics, Control, Staining, Immunohistochemistry

JAK2 is involved in a model of pulmonary fibrosis in mice constructed with bleomycin. (A) Expression of proteins associated with pulmonary fibrosis in mice by pirfenidone and nintedanib intervention. The protein expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 in mouse lung was measured by Western blotting. GAPDH was used as the internal control (n=6, ## , P<0.01; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with BLM group). (B) Expression of p-JAK2 in pathological sections of mouse lungs was detected by immunofluorescence staining. TGF, transforming growth factor; SMA, smooth muscle actin; DAPI, 4',6-diamidino-2-phenylindole; BLM, bleomycin.

Journal: Journal of Thoracic Disease

Article Title: Pirfenidone and nintedanib attenuate pulmonary fibrosis in mice by inhibiting the expression of JAK2

doi: 10.21037/jtd-23-1057

Figure Lengend Snippet: JAK2 is involved in a model of pulmonary fibrosis in mice constructed with bleomycin. (A) Expression of proteins associated with pulmonary fibrosis in mice by pirfenidone and nintedanib intervention. The protein expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 in mouse lung was measured by Western blotting. GAPDH was used as the internal control (n=6, ## , P<0.01; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with BLM group). (B) Expression of p-JAK2 in pathological sections of mouse lungs was detected by immunofluorescence staining. TGF, transforming growth factor; SMA, smooth muscle actin; DAPI, 4',6-diamidino-2-phenylindole; BLM, bleomycin.

Article Snippet: Anti-TGF-β-R2 antibodies were purchased from Proteintech.

Techniques: Construct, Expressing, Western Blot, Control, Immunofluorescence, Staining

In an in vitro model, JAK2 is involved in the signaling pathway of fibrosis. (A) The expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 proteins were detected by Western blotting after stimulation of MLE12 cell with different concentrations of TGF-β1 (0, 5, 10, 20, 50 and 100 ng/mL) for 48 h; GAPDH was used as the internal control (n=3, *, P<0.05; **, P<0.01; ***, P<0.001, compared with 0 ng/mL). (B) After treatment with TGF-β1 (50 ng/mL) for 6 h, the inhibitor of TGF-β1 receptor LY2109761 (0.01, 0.05 and 0.5 μM) was then applied to MLE12 at different concentrations, and Western blotting was performed to detect Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 protein expression levels. GAPDH was used as the internal control (n=3, ## , P<0.01; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with TGF-β1 group). (C) MLE12 was treated with TGF-β1 (50 ng/mL) for 6 h and then treated with NC and si-JAK2, and the expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 proteins were detected by Western blotting (n=3, ## , P<0.01; ### , P<0.001, compared with Control. **, P<0.01; ***, P<0.001, compared with TGF-β1 group). Fn, fibronectin; TGF, transforming growth factor; SMA, smooth muscle actin; NC, negative control.

Journal: Journal of Thoracic Disease

Article Title: Pirfenidone and nintedanib attenuate pulmonary fibrosis in mice by inhibiting the expression of JAK2

doi: 10.21037/jtd-23-1057

Figure Lengend Snippet: In an in vitro model, JAK2 is involved in the signaling pathway of fibrosis. (A) The expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 proteins were detected by Western blotting after stimulation of MLE12 cell with different concentrations of TGF-β1 (0, 5, 10, 20, 50 and 100 ng/mL) for 48 h; GAPDH was used as the internal control (n=3, *, P<0.05; **, P<0.01; ***, P<0.001, compared with 0 ng/mL). (B) After treatment with TGF-β1 (50 ng/mL) for 6 h, the inhibitor of TGF-β1 receptor LY2109761 (0.01, 0.05 and 0.5 μM) was then applied to MLE12 at different concentrations, and Western blotting was performed to detect Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 protein expression levels. GAPDH was used as the internal control (n=3, ## , P<0.01; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with TGF-β1 group). (C) MLE12 was treated with TGF-β1 (50 ng/mL) for 6 h and then treated with NC and si-JAK2, and the expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 proteins were detected by Western blotting (n=3, ## , P<0.01; ### , P<0.001, compared with Control. **, P<0.01; ***, P<0.001, compared with TGF-β1 group). Fn, fibronectin; TGF, transforming growth factor; SMA, smooth muscle actin; NC, negative control.

Article Snippet: Anti-TGF-β-R2 antibodies were purchased from Proteintech.

Techniques: In Vitro, Expressing, Western Blot, Control, Negative Control

In an in vitro model, pirfenidone and nintedanib also significantly attenuated indicators of pulmonary fibrosis. (A) MLE12 was treated with TGF-β1 (50 ng/mL) for 6 h and then intervened with pirfenidone (1 mM) and nintedanib (1 μM), and Western blotting was performed to detect Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 protein expression levels. GAPDH was used as the internal control (n=3, ## , P<0.01; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with TGF-β1 group). (B) TGF-β1 (50 ng/mL) was treated with MLE12 cell at different time points (0, 1, 2, 6, 12, 24 h), and the expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 proteins were detected by Western blotting. GAPDH was used as the internal control (n=3, ***, P<0.001, compared with 0 h). Fn, fibronectin; TGF, transforming growth factor; SMA, smooth muscle actin.

Journal: Journal of Thoracic Disease

Article Title: Pirfenidone and nintedanib attenuate pulmonary fibrosis in mice by inhibiting the expression of JAK2

doi: 10.21037/jtd-23-1057

Figure Lengend Snippet: In an in vitro model, pirfenidone and nintedanib also significantly attenuated indicators of pulmonary fibrosis. (A) MLE12 was treated with TGF-β1 (50 ng/mL) for 6 h and then intervened with pirfenidone (1 mM) and nintedanib (1 μM), and Western blotting was performed to detect Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 protein expression levels. GAPDH was used as the internal control (n=3, ## , P<0.01; ### , P<0.001, compared with Control. *, P<0.05; **, P<0.01; ***, P<0.001, compared with TGF-β1 group). (B) TGF-β1 (50 ng/mL) was treated with MLE12 cell at different time points (0, 1, 2, 6, 12, 24 h), and the expression levels of Fn, TGF-β-R2, α-SMA, p-JAK2, JAK2, p-STAT3, and STAT3 proteins were detected by Western blotting. GAPDH was used as the internal control (n=3, ***, P<0.001, compared with 0 h). Fn, fibronectin; TGF, transforming growth factor; SMA, smooth muscle actin.

Article Snippet: Anti-TGF-β-R2 antibodies were purchased from Proteintech.

Techniques: In Vitro, Western Blot, Expressing, Control

Silencing of MED12 increases the chemoresistance in colorectal cancer (CRC) cell lines. (A and B) CRC cells were transfected with either scrambled siRNA or MED12- specific small interfering RNA (siRNA) for 48 hours and clonogenic assay were performed. Cells (6×10 2 ) after post-transfection were seeded into each of three dishes (60 mm diameter), and grown for an additional 10 days, then stained with crystal violet (A). Colony numbers in the entire dish were counted (B). (C) CRC cells were transfected with either scrambled siRNA or MED12 siRNA and subsequently treated with 50 and 100 μM 5-fluorouracil (FU) for 48 hours. Following treatment, cells were analysed for apoptosis by flow cytometry. (D) LOVO and DLD1 cells were transfected with either scrambled siRNA or MED12- specific siRNA for 48 hours. Proteins were isolated and immunoblotted with antibodies against MED12, transforming growth factor (TGF)-β-R2, p-ERK1/2, ERK1/2, E-cadherin, N-cadherin, vimentin, Twist and β-actin for equal loading. (E) Representative images of fluorescence immunostaining for MED12, TGF-β-R2 and E-cadherin in LOVO and DLD1 cells after post-transfection with MED12 siRNA. Data presented in the bar graphs are the mean±SD of three independent experiments. *Indicates a statistically significant difference compared with control with p<0.05.

Journal: Gut

Article Title: MED12 is recurrently mutated in Middle Eastern colorectal cancer

doi: 10.1136/gutjnl-2016-313334

Figure Lengend Snippet: Silencing of MED12 increases the chemoresistance in colorectal cancer (CRC) cell lines. (A and B) CRC cells were transfected with either scrambled siRNA or MED12- specific small interfering RNA (siRNA) for 48 hours and clonogenic assay were performed. Cells (6×10 2 ) after post-transfection were seeded into each of three dishes (60 mm diameter), and grown for an additional 10 days, then stained with crystal violet (A). Colony numbers in the entire dish were counted (B). (C) CRC cells were transfected with either scrambled siRNA or MED12 siRNA and subsequently treated with 50 and 100 μM 5-fluorouracil (FU) for 48 hours. Following treatment, cells were analysed for apoptosis by flow cytometry. (D) LOVO and DLD1 cells were transfected with either scrambled siRNA or MED12- specific siRNA for 48 hours. Proteins were isolated and immunoblotted with antibodies against MED12, transforming growth factor (TGF)-β-R2, p-ERK1/2, ERK1/2, E-cadherin, N-cadherin, vimentin, Twist and β-actin for equal loading. (E) Representative images of fluorescence immunostaining for MED12, TGF-β-R2 and E-cadherin in LOVO and DLD1 cells after post-transfection with MED12 siRNA. Data presented in the bar graphs are the mean±SD of three independent experiments. *Indicates a statistically significant difference compared with control with p<0.05.

Article Snippet: Antibodies against transforming growth factor (TGF)-β-R2 (sc#17799), vimentin (sc#5565) and GAPDH (sc#25778) were purchased from Santa Cruz Biotechnology (Santa Cruz, California, USA).

Techniques: Transfection, Small Interfering RNA, Clonogenic Assay, Staining, Flow Cytometry, Isolation, Fluorescence, Immunostaining, Control

Forced expression of MED12 decreases the chemoresistance in colorectal cancer (CRC) cell lines. (A and B) CRC cells were transfected with either empty vector or MED12 pcDNA for 48 hours and clonogenic assay were performed. Cells (6×10 2 ) after post-transfection were seeded into each of three dishes (60 mm diameter), and grown for an additional 10 days, then stained with crystal violet (A). Colony numbers in the entire dish were counted (B). (C) CRC cells were transfected with either empty vector or MED12 pcDNA and subsequently treated with 50 and 100 μM 5-fluorouracil (FU) for 48 hours. Following treatment, cells were analysed for apoptosis by flow cytometry. (D) COLO-320 and HT29 cells were transfected with either empty vector or MED12 pcDNA for 48 hours. Proteins were isolated and immunoblotted with antibodies against MED12, transforming growth factor (TGF)-β-R2, p-ERK1/2, ERK1/2, E-cadherin, N-cadherin, vimentin, Twist and GAPDH for equal loading. (E) Representative images of fluorescence immunostaining for MED12, TGF-β-R2 and E-cadherin in COLO-320 and HT29 cells after post-transfection with MED12 pcDNA. Data presented in the bar graphs are the mean±SD of three independent experiments. *Indicates a statistically significant difference compared with control with p<0.05.

Journal: Gut

Article Title: MED12 is recurrently mutated in Middle Eastern colorectal cancer

doi: 10.1136/gutjnl-2016-313334

Figure Lengend Snippet: Forced expression of MED12 decreases the chemoresistance in colorectal cancer (CRC) cell lines. (A and B) CRC cells were transfected with either empty vector or MED12 pcDNA for 48 hours and clonogenic assay were performed. Cells (6×10 2 ) after post-transfection were seeded into each of three dishes (60 mm diameter), and grown for an additional 10 days, then stained with crystal violet (A). Colony numbers in the entire dish were counted (B). (C) CRC cells were transfected with either empty vector or MED12 pcDNA and subsequently treated with 50 and 100 μM 5-fluorouracil (FU) for 48 hours. Following treatment, cells were analysed for apoptosis by flow cytometry. (D) COLO-320 and HT29 cells were transfected with either empty vector or MED12 pcDNA for 48 hours. Proteins were isolated and immunoblotted with antibodies against MED12, transforming growth factor (TGF)-β-R2, p-ERK1/2, ERK1/2, E-cadherin, N-cadherin, vimentin, Twist and GAPDH for equal loading. (E) Representative images of fluorescence immunostaining for MED12, TGF-β-R2 and E-cadherin in COLO-320 and HT29 cells after post-transfection with MED12 pcDNA. Data presented in the bar graphs are the mean±SD of three independent experiments. *Indicates a statistically significant difference compared with control with p<0.05.

Techniques: Expressing, Transfection, Plasmid Preparation, Clonogenic Assay, Staining, Flow Cytometry, Isolation, Fluorescence, Immunostaining, Control

TGF- β signaling is essential for drug resistance induced by ZNF32 overexpression. ( a ) Western blot analysis of MEK/ERK signaling in PC9 cells in both the absence and presence of GEF (10 μ M). ( b ) qRT-PCR detection of TGF- β R2 and TGF- β target gene (CDH2, TAGLN and CYR61) expression in A549 and PC9 cells. ( c ) Western blot analysis of TGF- β R2 expression and SMAD2 (pSMAD2) phosphorylation in PC9 cells. ( d ) In PC9 cells, the combination of ZNF32 overexpression and recombinant TGF- β (10 ng/ml) activates TGF- β and MEK/ERK signaling, and LY2157299 (1 μ M) inhibits TGF- β and the majority of MEK/ERK signaling. ( e ) and ( f ) A 3D colony-forming assay and a flow cytometric analysis confirm that TGF- β can induce resistance in AC cells, whereas LY2157299 can counteract the effect of ZNF32 and cancel this resistance. NS, non-significant difference. Each column and bar represents the mean±S.D. of three independent experiments. The photograph shows a representative result from three independent experiments

Journal: Cell Death & Disease

Article Title: ZNF32 contributes to the induction of multidrug resistance by regulating TGF- β receptor 2 signaling in lung adenocarcinoma

doi: 10.1038/cddis.2016.328

Figure Lengend Snippet: TGF- β signaling is essential for drug resistance induced by ZNF32 overexpression. ( a ) Western blot analysis of MEK/ERK signaling in PC9 cells in both the absence and presence of GEF (10 μ M). ( b ) qRT-PCR detection of TGF- β R2 and TGF- β target gene (CDH2, TAGLN and CYR61) expression in A549 and PC9 cells. ( c ) Western blot analysis of TGF- β R2 expression and SMAD2 (pSMAD2) phosphorylation in PC9 cells. ( d ) In PC9 cells, the combination of ZNF32 overexpression and recombinant TGF- β (10 ng/ml) activates TGF- β and MEK/ERK signaling, and LY2157299 (1 μ M) inhibits TGF- β and the majority of MEK/ERK signaling. ( e ) and ( f ) A 3D colony-forming assay and a flow cytometric analysis confirm that TGF- β can induce resistance in AC cells, whereas LY2157299 can counteract the effect of ZNF32 and cancel this resistance. NS, non-significant difference. Each column and bar represents the mean±S.D. of three independent experiments. The photograph shows a representative result from three independent experiments

Article Snippet: The antibodies used in this assay were the following: mouse anti-ZNF32 antibody (1:200), mouse anti-TGF- β R2 antibody (1:100), rabbit anti- β -actin antibody (1:800, Santa Cruz Biotechnology), rabbit anti-ERK antibody (1:1000, Cell Signaling Technology, Lexington, KY, USA), rabbit anti-p-ERK antibody (1:500, Cell Signaling Technology), rabbit anti-SMAD2 antibody (1:500, Cell Signaling Technology), rabbit anti-pSMAD2 antibody (1:200, Cell Signaling Technology), rabbit anti-MEK antibody (1:1000, Cell Signaling Technology), rabbit anti-p-MEK antibody (1:500, Cell Signaling Technology), horseradish peroxidase-conjugated secondary antibody to rabbit IgG (1:5000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and horseradish peroxidase-conjugated secondary antibody to mouse IgG (1:8000, Santa Cruz Biotechnology).

Techniques: Over Expression, Western Blot, Quantitative RT-PCR, Expressing, Phospho-proteomics, Recombinant

The transcription of TGF- β R2 is regulated by ZNF32. ( a ) An immunofluorescence assay shows stronger TGF- β R2 (green) expression in the Lv-ZNF32 group and weaker expression in the Sh-ZNF32 group. ( b ) Schematic representation of the ZNF32-binding sites in the TGF- β R2 promoter. The probe sequence is underlined, and the transcription start site is indicated by +1. ( c ) HEK293 cells were transiently transfected with TGF- β R2 promoter 5′-deletion mutant constructs and analyzed through a dual-luciferase reporter assay. ( d ) HEK293 cells were transiently transfected with the indicated constructs, treated as in ( c ) and then analyzed using a dual-luciferase reporter assay. ( e ) Nuclear extracts from A549 cells were incubated in biotin-labeled oligonucleotides corresponding to the TGF- β R2 promoter region −736/−751. The arrow shows the specific DNA-protein complex. ( f ) DNA fragments from A549 cells were immunoprecipitated with ZNF32-specific antibodies and analyzed via RT-PCR using the indicated primers. NS, non-significant difference. The data are presented as the means±S.D. Each experiment was performed at least in triplicate, and consistent results were obtained. The photograph shows a representative result from three independent experiments

Journal: Cell Death & Disease

Article Title: ZNF32 contributes to the induction of multidrug resistance by regulating TGF- β receptor 2 signaling in lung adenocarcinoma

doi: 10.1038/cddis.2016.328

Figure Lengend Snippet: The transcription of TGF- β R2 is regulated by ZNF32. ( a ) An immunofluorescence assay shows stronger TGF- β R2 (green) expression in the Lv-ZNF32 group and weaker expression in the Sh-ZNF32 group. ( b ) Schematic representation of the ZNF32-binding sites in the TGF- β R2 promoter. The probe sequence is underlined, and the transcription start site is indicated by +1. ( c ) HEK293 cells were transiently transfected with TGF- β R2 promoter 5′-deletion mutant constructs and analyzed through a dual-luciferase reporter assay. ( d ) HEK293 cells were transiently transfected with the indicated constructs, treated as in ( c ) and then analyzed using a dual-luciferase reporter assay. ( e ) Nuclear extracts from A549 cells were incubated in biotin-labeled oligonucleotides corresponding to the TGF- β R2 promoter region −736/−751. The arrow shows the specific DNA-protein complex. ( f ) DNA fragments from A549 cells were immunoprecipitated with ZNF32-specific antibodies and analyzed via RT-PCR using the indicated primers. NS, non-significant difference. The data are presented as the means±S.D. Each experiment was performed at least in triplicate, and consistent results were obtained. The photograph shows a representative result from three independent experiments

Techniques: Immunofluorescence, Expressing, Binding Assay, Sequencing, Transfection, Mutagenesis, Construct, Luciferase, Reporter Assay, Incubation, Labeling, Immunoprecipitation, Reverse Transcription Polymerase Chain Reaction

ZNF32 deficiency might exhibit synergistic effects with a TGF- β R inhibitor to augment the anti-tumor effect of drugs and improve patient survival in vivo . Twenty-nine days after the injection of A549 and PC9 cells into the mice, the tumor mass was obtained. ( a ) Volume and weight of the tumor. ( b ) Growth curve of the tumor. ( c ) These samples were sliced and stained with HE to measure the necrosis area. ( d ) When the mice died, the survival time of each group was recorded, and the Kaplan–Meier survival curves for each group were analyzed ( n =10 per group; * P <0.05). NS, non-significant difference. Each column and bar represents the median±S.D. of three independent experiments. The photograph shows a representative result from three independent experiments

Journal: Cell Death & Disease

Article Title: ZNF32 contributes to the induction of multidrug resistance by regulating TGF- β receptor 2 signaling in lung adenocarcinoma

doi: 10.1038/cddis.2016.328

Figure Lengend Snippet: ZNF32 deficiency might exhibit synergistic effects with a TGF- β R inhibitor to augment the anti-tumor effect of drugs and improve patient survival in vivo . Twenty-nine days after the injection of A549 and PC9 cells into the mice, the tumor mass was obtained. ( a ) Volume and weight of the tumor. ( b ) Growth curve of the tumor. ( c ) These samples were sliced and stained with HE to measure the necrosis area. ( d ) When the mice died, the survival time of each group was recorded, and the Kaplan–Meier survival curves for each group were analyzed ( n =10 per group; * P <0.05). NS, non-significant difference. Each column and bar represents the median±S.D. of three independent experiments. The photograph shows a representative result from three independent experiments

Techniques: In Vivo, Injection, Staining

ZNF32 is positively correlated with TGF- β R2 expression and negatively correlated with prognosis. ( a ) IHC detection of the expression of ZNF32 and its downstream target TGF- β R2 in AC samples (left panel). A correlation analysis demonstrates that ZNF32 expression is positively correlated with TGF- β R2 expression in AC tissues (right panel). ( b ) Kaplan–Meier survival curve for 52 patients who were categorized into two groups based on the nuclear ZNF32 IHC score. ( c ) Kaplan–Meier survival curve for 37 patients who had received CIS-based chemotherapy, whose responses to chemotherapy are known, and who were categorized into two groups based on the nuclear ZNF32 IHC score. ( d ) Multivariate analysis of ZNF32 expression in patients. The risk ratio (proportional hazard) was calculated with respect to the following parameters: ±nuclear ZNF32 expression, stage (I/II–IIIA), grade (high/low), Ki67 (high/low), and sex (male/female). ( e ) Comparison of ZNF32 expression between patients with and without brain metastasis. ( f ) Tissue slices were cultured with CIS (20 μ M) for 3 days, and IHC was then performed for the comparison of Ki-67 expression and TUNEL positivity between the ZNF32 high and ZNF32 low groups ( n =5 per group). NS, non-significant difference. Each column and bar represents the median±S.D. of three independent experiments. The photograph shows a representative result from three independent experiments

Journal: Cell Death & Disease

Article Title: ZNF32 contributes to the induction of multidrug resistance by regulating TGF- β receptor 2 signaling in lung adenocarcinoma

doi: 10.1038/cddis.2016.328

Figure Lengend Snippet: ZNF32 is positively correlated with TGF- β R2 expression and negatively correlated with prognosis. ( a ) IHC detection of the expression of ZNF32 and its downstream target TGF- β R2 in AC samples (left panel). A correlation analysis demonstrates that ZNF32 expression is positively correlated with TGF- β R2 expression in AC tissues (right panel). ( b ) Kaplan–Meier survival curve for 52 patients who were categorized into two groups based on the nuclear ZNF32 IHC score. ( c ) Kaplan–Meier survival curve for 37 patients who had received CIS-based chemotherapy, whose responses to chemotherapy are known, and who were categorized into two groups based on the nuclear ZNF32 IHC score. ( d ) Multivariate analysis of ZNF32 expression in patients. The risk ratio (proportional hazard) was calculated with respect to the following parameters: ±nuclear ZNF32 expression, stage (I/II–IIIA), grade (high/low), Ki67 (high/low), and sex (male/female). ( e ) Comparison of ZNF32 expression between patients with and without brain metastasis. ( f ) Tissue slices were cultured with CIS (20 μ M) for 3 days, and IHC was then performed for the comparison of Ki-67 expression and TUNEL positivity between the ZNF32 high and ZNF32 low groups ( n =5 per group). NS, non-significant difference. Each column and bar represents the median±S.D. of three independent experiments. The photograph shows a representative result from three independent experiments

Techniques: Expressing, Comparison, Cell Culture, TUNEL Assay

Loss of Tfrc in NCCs suppresses the activation of TGF- β and BMP signaling. ( a ) Expression levels of indicated signaling pathways targeted genes in mandible tissues dissected from E13.5 Wnt1 cre ;Tfrc f/f mutants and controls. Data shown are normalized ratio of Wnt1 cre ;Tfrc f/f /Control (mean±S.E.M); student's t -test; NS, no significant difference; *** P <0.001; n ≥3. ( b ) Expression levels of proteins involved in TGF- β and BMP signaling in E13.5 mandible tissues. Data shown are normalized ratio of Wnt1 cre ;Tfrc f/f /Control (mean±S.E.M); student's t -test; * P <0.05; ** P <0.01; n ≥3. ( c ) Immunostaining of P-Smad5 in E13.5 mandible sections. P, palate; T, tongue; Mc, Meckel's cartilage. Scale bar, 100 μ m. ( d and e ) Immunoblotting and quantification of P-smad5 ( c ) and P-smad2 ( d ) in cultured primary mandibular mesenchymal cells. Mandibular mesenchymal cells were cultured and induced to undergo osteochondrogenic differentiation for 7 days, then treated with BMP2 (100 ng/ml) or Tgf β 3 (50 ng/ml) for 30 min at 37 °C. Data shown are normalized ratio of KO/Control (mean±S.E.M); student's t -test; * P <0.05; ** P <0.01; n =3

Journal: Cell Death & Disease

Article Title: Transferrin receptor facilitates TGF- β and BMP signaling activation to control craniofacial morphogenesis

doi: 10.1038/cddis.2016.170

Figure Lengend Snippet: Loss of Tfrc in NCCs suppresses the activation of TGF- β and BMP signaling. ( a ) Expression levels of indicated signaling pathways targeted genes in mandible tissues dissected from E13.5 Wnt1 cre ;Tfrc f/f mutants and controls. Data shown are normalized ratio of Wnt1 cre ;Tfrc f/f /Control (mean±S.E.M); student's t -test; NS, no significant difference; *** P <0.001; n ≥3. ( b ) Expression levels of proteins involved in TGF- β and BMP signaling in E13.5 mandible tissues. Data shown are normalized ratio of Wnt1 cre ;Tfrc f/f /Control (mean±S.E.M); student's t -test; * P <0.05; ** P <0.01; n ≥3. ( c ) Immunostaining of P-Smad5 in E13.5 mandible sections. P, palate; T, tongue; Mc, Meckel's cartilage. Scale bar, 100 μ m. ( d and e ) Immunoblotting and quantification of P-smad5 ( c ) and P-smad2 ( d ) in cultured primary mandibular mesenchymal cells. Mandibular mesenchymal cells were cultured and induced to undergo osteochondrogenic differentiation for 7 days, then treated with BMP2 (100 ng/ml) or Tgf β 3 (50 ng/ml) for 30 min at 37 °C. Data shown are normalized ratio of KO/Control (mean±S.E.M); student's t -test; * P <0.05; ** P <0.01; n =3

Article Snippet: Antibodies used in immunoblotting analysis: Tfrc (Invitrogen, Waltham, MA, USA, 136800), Phospho-Smad5 (Epitomics, Burlingame, CA, USA, 2224-1), Phospho-Smad2 (Cell Signaling Technology, 3101), BMPR2 (Epitomics, S0778), Tgf β R2 (Cell Signaling Technology, Burlingame, CA, USA, 3713), Runx2 (Millipore, Merck, KGaA, Darmstadt, Germany, 05-1478), β -actin (Santa Cruz, sc-47778).

Techniques: Activation Assay, Expressing, Protein-Protein interactions, Control, Immunostaining, Western Blot, Cell Culture

Review

tgf β r2 (Santa Cruz Biotechnology)

Structured Review

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1) Product Images from "CKD-Induced Oxidative Twitch Muscle Atrophy Is Mediated by TGF- β"

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