cct2 (Santa Cruz Biotechnology)
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Structured Review
Santa Cruz Biotechnology
cct2
Cct2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cct2/TCP-1+%CE%B2+Antibody/pmc12718141-257-83-86
Average 93 stars, based on 13 article reviews
Cct2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cct2/TCP-1+%CE%B2+Antibody/pmc12718141-257-83-86
Average 93 stars, based on 13 article reviews
cct2 - by Bioz Stars,
2026-09
93/100 stars
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1) Product Images from "Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis"
Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis
Journal: The Journal of Biological Chemistry
doi: 10.1016/j.jbc.2025.110914
Figure Legend Snippet: FKBP4 prevents CCT8 aggregation. A , following the knockdown of FKBP4 in HCT116 cells for 72 h, the aggregation of CCT1 and CCT8 proteins within cells was assessed using a filter trap assay. Cell lysates were filtered through a nitrocellulose membrane, and the retained proteins on the membrane were detected using specific antibodies against CCT1 and CCT8. B , quantitative analysis of CCT1 and CCT8 protein aggregation, normalized to GAPDH, in shFKBP4 HCT116 cells. C , cell lysates that did not pass through the nitrocellulose membrane were subjected to western blotting to detect the levels of FKBP4, CCT8, CCT1, CDK2, α-tubulin, and GAPDH within the cells. D , quantitative analysis of CCT8 and CCT1 protein expression, normalized to GAPDH, in shFKBP4 HCT116 cells. E , following the knockdown of FKBP4 in HCT116 cells for 72 h, the aggregation of CCT2 and CCT3 proteins within cells was assessed using a filter trap assay. The experimental procedure mirrored that of ( A ). F , quantitative analysis of CCT2 and CCT3 protein aggregation, normalized to GAPDH, in shFKBP4 HCT116 cells. G , cell lysates that did not pass through the nitrocellulose membrane were subjected to western blotting to detect the levels of FKBP4, CCT2, CCT3, and GAPDH within the cells. The experimental procedure mirrored that of ( C ). H , quantitative analysis of CCT2 and CCT3 protein expression, normalized to GAPDH, in shFKBP4 HCT116 cells. I , shFKBP4 HCT116 cells were harvested after 3 days of puromycin selection. The cell lysates were biochemically fractionated into Triton-soluble and -insoluble fractions as described in “ ”. The FKBP4 expression levels and Triton-soluble and -insoluble fractions of CCT8 were analyzed by western blotting. The GAPDH was used as a loading control. J , The ratio of the CCT8 Triton-insoluble form in ( I ). K , The primary structures of FKBP4 and FKBP5. L , after 48 h of transfection, vector, FLAG-tagged FKBP4 or FLAG-tagged FKBP5 expressing HEK293T cell lysates were precipitated by anti-FLAG antibodies, and the products were detected for the co-purification of the endogenous proteins. M , comparison of the interaction between Hsp90 and FLAG-tagged FKBP4 or FLAG-tagged FKBP5 in ( L ). N , following the knockdown of FKBP5 in HCT116 cells for 72 h, the aggregation of CCT8 proteins within cells was assessed using a filter trap assay. The experimental procedure mirrored that of ( A ). O , quantitative analysis of the protein aggregation of CCT8, normalized to GAPDH, in shFKBP5 HCT116 cells. P , cell lysates that did not pass through the nitrocellulose membrane were subjected to western blotting to detect the levels of FKBP5, CCT8, and GAPDH within the cells. The experimental procedure mirrored that of ( C ). Q , quantitative analysis of CCT8 protein expression, normalized to GAPDH, in shFKBP5 HCT116 cells. R , shFKBP5 HCT116 cells were harvested after puromycin selection for 3 days. The experimental procedure mirrored that of ( I ). S , the ratio of the CCT8 Triton-insoluble form in ( R ). T , HCT116 cells were treated with 17-AAG (20 μM) or ganetespib (2 μM) for 24 h, and CCT8 aggregation was assessed by filter trap assay. U , quantitative analysis of CCT8 protein aggregation with GAPDH serving as an internal control in ( T ). V , cell lysates that did not pass through the nitrocellulose membrane were subjected to western blotting to detect the levels of HSPA1A, CCT8, HSP90, and GAPDH within the cells. The experimental procedure mirrored that of ( C ). W , quantitative analysis of CCT8 protein expression was normalized with GAPDH in ( V ). All data were analyzed with an unpaired two-tailed Student's t test. Each dataset is expressed as mean ± SD for n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.
Techniques Used: Knockdown, TRAP Assay, Membrane, Western Blot, Expressing, Selection, Control, Transfection, Plasmid Preparation, Copurification, Comparison, Two Tailed Test
Related Articles
Knockdown:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from TRAP Assay:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Membrane:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Western Blot:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Expressing:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Selection:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Control:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Transfection:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Plasmid Preparation:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Copurification:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Comparison:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Two Tailed Test:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Recombinant:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Staining:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Negative Control:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Software:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from SDS Page:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Over Expression:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from Labeling:Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam), GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD), FASN (1:1000, GTX109833, GeneTex), FLNA (1:1000, GTX112939, GeneTex), TALIN-1 (1:500, GTX102215, GeneTex), MRE11 (1:1000, PC388, Calbiochem), DDX3 (1:1000, GTX110614, GeneTex), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex), CDK2 (1:500, 2546, Cell Signaling), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology), FKBP5 (1:1000, GTX113438, GeneTex), α-tubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc), FLAG (1:2000, F3165, Sigma-Aldrich), c-Myc (1:1000, #11667203001, Roche), CCT2 ( Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis. Article Snippet: Primary antibodies were used to detect FKBP4 (1:1000, ab97306, Abcam, USA), Jo urn al Pr e-p roo f 23 GAPDH (1:1000, GTX100118, GeneTex, USA), E-cadherin (1:1000, 610182, BD, USA), FASN (1:1000, GTX109833, GeneTex, USA), FLNA (1:1000, GTX112939, GeneTex, USA), TALIN-1 (1:500, GTX102215, GeneTex, USA), MRE11 (1:1000, PC388, Calbiochem, USA), DDX3 (1:1000, GTX110614, GeneTex, USA), TCP1 theta (CCT8) (1:1000, GTX105725, GeneTex, USA), CDK2 (1:500, 2546, Cell Signaling, USA), CCT1 (1:250, sc-53454, Santa-cruz Biotechnology, USA), HSP90 (1:1000, sc-13119, Santa-cruz Biotechnology, USA), HSPA1A (1:1000, sc-66048, Santa-cruz Biotechnology, USA), FKBP5 (1:1000, GTX113438, GeneTex, USA), αtubulin (1:1000, MCA78G, Bio-Rad Laboratories Inc., USA), FLAG (1:2000, F3165, Sigma-Aldrich, USA), c-Myc (1:1000, 11667203001, Roche, Switzerland), CCT2 ( Article Title: Mechanical control of the alternative splicing factor PTBP1 regulates extracellular matrix stiffness induced proliferation and cell spreading Article Snippet: The membranes were blocked in blocking buffer (5% skim milk, 0.05% Tween 20, 1xPBS) for 1 h, followed by incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies.incubation overnight at 4°C with primary antibodies diluted in blocking buffer and then 1 h at room temperature with secondary antibodies. ... Primary antibodies were used as follows: YAP (1:200; Santa Cruz, sc-101199), GFP (1:2000; Invitrogen, A-11122), PTBP1 (1:500; Thermo Fisher Scientific, 32–4800), Pitx2 (1:1000; Capra Science, PA-1020), Article Title: Mechanical control of the splicing factor PTBP1 regulates extracellular matrix stiffness-induced cell proliferation and mechanomemory Article Snippet: The membranes were blocked in 5% (w/v) milk in PBST (0.05% Tween 20, 1xPBS) for 1 hour, followed by incubation overnight at 4°C with primary antibodies diluted in 5% (w/v) milk in PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies.PBST or in 5% (w/v) BSA in TBST (13.7 mM NaCl, 2 mM Tris-HCl, 0.1% Tween 20 pH 7.6) and then 1 hour at room temperature with secondary antibodies. ... Primary antibodies were used at the following dilutions: YAP 1:200 (sc-101199; Santa Cruz), GFP 1:2000 (A-11122; Invitrogen), PTBP1 1:500 (32-4800; Thermo Fisher Scientific), Pitx2 1:1000 (PA-1020; Capra Science), Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification. Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: THOC3 interacts with YBX1 to promote lung squamous cell carcinoma progression through PFKFB4 mRNA modification Article Snippet: The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).The membranes were then incubated with primary antibodies, followed by secondary antibodies, and protein levels were detected using an enhanced chemiluminescence reagent (xinsaimei, China).. Antibodies were purchased from manufacturers: THOC3 (Santa Cruz, USA, sc-377456), YBX1 (Proteintech, China, 20339-1-AP), Flag (CST, USA, #14793), β-Actin (CST, USA, #3700), PFKFB4 (Bioss, China, bs-12632R), NSUN2(Bioss, China, bs-19477R), CCT8 (Santa Cruz, USA, sc-377261), CCT6A(Santa Cruz, USA, sc-514466), and Article Title: Co-expression of CCT subunits hints at TRiC assembly Article Snippet: Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).Alternatively, transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 mM Tris, 192 mM glycine) onto 0.45 μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies were from Article Title: Human CCT4 and CCT5 Chaperonin Subunits Expressed in Escherichia coli Form Biologically Active Homo-oligomers Article Snippet: Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).Transfer was conducted for 1.5 h at 300 mA in transfer buffer (10% methanol, 25 m m Tris, 192 m m glycine) onto 0.45-μm polyvinylidene difluoride (PVDF) membranes (Millipore).. The primary antibodies used for CCT1–8 were from |
