cct2  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: Stratifying cellular injury in Alzheimer’s disease by chaperonin containing TCP1 subunits 2 and 3

doi: 10.64898/2026.05.10.724132

Figure Lengend Snippet: ( a ) In the dentate gyrus, CCT2 accumulated in granule cells, pyramidal cells of the CA4 sub-field, and hilar neurons. ( a 1 ) CCT2-like immunoreactivity was reduced as the granule cell layer became disorganized in definite AD. ( b ) CCT2 in CA1 pyramidal neurons, particularly in their somata and proximal dendrites. ( b 1 ) Here, CCT2-like immunoreactivity was also reduced in definite AD. ( c ) In AD, an increased number of tangle-bearing neurons and extracellular plaques ( arrowheads ) were associated with reduced CCT3-like immunoreactivity in neurons in, e.g., the polymorph layer of the dentate gyrus (PoDG; c 1 ). In definite AD ( d 1 vs . d ), perisomatic CCT3-like immunoreactivity was absent in those CA1 pyramidal neurons that were proximal to extracellular plaques ( arrowheads ). ( e,f ) Western blotting revealed gradually reduction in both CCT2 and CCT3 protein levels as a factor of disease progression in the human hippocampus. For CCT3, two isoforms (‘a’ (75 kDa) ‘b’ (65 kDa), corresponding to possible splice variants) were quantified. β-Actin was used as a house-keeping standard. Representative Western blots were shown (e 1 ,f 1 ) with a total of n = 21 cases quantified. # p < 0.1, * p < 0.05, ** p <0.001. Abbreviations : cont, control; def, definite AD; GL, granule cell layer; mol, molecular layer; pos, possible AD; prob, probable AD; pyr, pyramidal layer; rad, radial layer. Scale bars = 40 µm (a 1 ,c 1 ), 15 µm (b 1 ,d 1 ).

Article Snippet: Antibody generation and specificity ( a ) Within the Human Protein Atlas (HPA) project, rabbits were immunized against the amino acid sequences highlighted within CCT2 (2x) and CCT3. ( b ) The specificity of anti-CCT2 antibodies (note their HPA database designations) were first tested in protein Epitope Tag (PrEST) screens. ( c ) Subsequently, antibody specificity was substantiated on Western blots from both cell lines and post-mortem hippocampal homogenates.

Techniques: Western Blot, Biomarker Discovery, Control

Journal: bioRxiv

Article Title: Stratifying cellular injury in Alzheimer’s disease by chaperonin containing TCP1 subunits 2 and 3

doi: 10.64898/2026.05.10.724132

Figure Lengend Snippet: ( a,b ) Intracellular Aβ accumulation in CA1 pyramidal neurons was found as early as in possible AD. ( c ) CCT2-like immunoreactivity was mutually exclusive with Aβ intracellularly; as was confirmed by plotting fluorescence signal intensity in affected neurons (along the line between 1,2 (example); d ). ( e ) Similarly, CCT3-like immunoreactivity was excluded from Aβ-laden sub-cellular areas ( f ). Asterisks denote the location of nuclei. Scale bars = 15 µm (a,b), 2 µm (e,f).

Article Snippet: Antibody generation and specificity ( a ) Within the Human Protein Atlas (HPA) project, rabbits were immunized against the amino acid sequences highlighted within CCT2 (2x) and CCT3. ( b ) The specificity of anti-CCT2 antibodies (note their HPA database designations) were first tested in protein Epitope Tag (PrEST) screens. ( c ) Subsequently, antibody specificity was substantiated on Western blots from both cell lines and post-mortem hippocampal homogenates.

Techniques: Fluorescence

Journal: bioRxiv

Article Title: Stratifying cellular injury in Alzheimer’s disease by chaperonin containing TCP1 subunits 2 and 3

doi: 10.64898/2026.05.10.724132

Figure Lengend Snippet: ( a ) Overview of the hippocampal formation and the parahippocampal gyrus immunostained for CCT2 from a subject with possible AD. ( b ) Heat-map of tau immunoreactivity color-coded from low (blue) to high (red) abundance. Arrowheads point to a tau-bearing locus. #1 and #2 are shown at higher magnification in panel (e). ( c,d ) The accumulation of hyperphosphorylated tau commenced in the perinuclear cytoplasm, with a gradual spread towards dendrites. Both CCT2-like and CCT3-like immunoreactivities were reduced subcellularly where tau accumulation occurred ( arrows ). Asterisks denote the positions of nuclei. ( e ) Comparison of CCT2-like immunoreactivity in cell ensembles in hippocampal CA1 in control vs . possible AD. ( f ) CCT2-like immunoreactivity was significantly reduced (* p <. 0.05). Scale bars = 100 µm (a), 50 µm (b), 17 µm (e), 2 µm (c,d).

Article Snippet: Antibody generation and specificity ( a ) Within the Human Protein Atlas (HPA) project, rabbits were immunized against the amino acid sequences highlighted within CCT2 (2x) and CCT3. ( b ) The specificity of anti-CCT2 antibodies (note their HPA database designations) were first tested in protein Epitope Tag (PrEST) screens. ( c ) Subsequently, antibody specificity was substantiated on Western blots from both cell lines and post-mortem hippocampal homogenates.

Techniques: Comparison, Control

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: In silico analysis of CCT2 expression and the clinical significance in HCC. Transcription levels of CCT2 in (A) TCGA-LIHC, (B) ICGC-LIRI-JP and (C) NODE-OEP00000321 datasets. ***P<0.001 vs. tumor-adjacent. Association between CCT2 expression and the overall survival prognosis of patients in (D) TCGA-LIHC, (E) ICGC-LIRI-JP and (F) NODE-OEP00000321 datasets. CCT2, chaperonin containing TCP1 subunit 2; TCGA-LIHC, The Cancer Genome Atlas-liver hepatocellular carcinoma; ICGC-LIRI-JP, International Cancer Genome Consortium-liver Cancer-Riken, Japan; NODE, National Omics Data Encyclopedia; HCC, hepatocellular carcinoma.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: In Silico, Expressing

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 in HCC cells. (A) Protein expression levels of CCT2 in HCC cell lines (Huh-7, Hep3B, Li-7 and HCCLM3). CCT2 was successfully knocked down in both Huh-7 and HCCLM3 cells following sh-CCT2 transduction. Knockdown efficiency in Huh-7 cells was evaluated by (B) reverse transcription-quantitative PCR and (C) western blotting. Knockdown efficiency in HCCLM3 cells was evaluated by (D) reverse transcription-quantitative PCR and (E) western blotting. *P<0.05, ***P<0.001 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; HCC, hepatocellular carcinoma.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Expressing, Transduction, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Negative Control

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits the proliferation and promotes the apoptosis of hepatocellular carcinoma cells. EdU incorporation assay was used to determine the DNA replication activity of (A) Huh-7 and (B) HCCLM3 cells. Colony formation assay was used to assess the cell clonogenicity of (C) Huh-7 and (D) HCCLM3 cells. Flow cytometry assay was used to analyze the percentage of apoptotic (E) Huh-7 and (F) HCCLM3 cells. Caspase-3/7 activity assay was used to evaluate the degree of apoptosis in (G) Huh-7 and (H) HCCLM3 cells. *P<0.05, **P<0.01, ***P<0.001 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; RFU, relative fluorescence units.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Activity Assay, Colony Assay, Flow Cytometry, Negative Control, Fluorescence

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits the migration and invasion of hepatocellular carcinoma cells. The effect of CCT2 knockdown on the migration of (A) Huh-7 and (B) HCCLM3 cells was measured using gap closure assay. The effects of CCT2 knockdown on the invasion of (C) Huh-7 and (D) HCCLM3 cells was evaluated by Transwell assay. *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Migration, Transwell Assay, Negative Control

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits the stemness characteristics of hepatocellular carcinoma cells. Tumor-sphere formation assay was used to assess the stem cell-like properties of (A) Huh-7 and (B) HCCLM3 cells following CCT2 knockdown. EdU incorporation assay was used to determine the proliferation of spheroid and parental (C) Huh-7 and (D) HCCLM3 cells following CCT2 knockdown. Transwell assay were used to evaluate the invasion of spheroid and parental (E) Huh-7 and (F) HCCLM3 cells following CCT2 knockdown. *P<0.05, **P<0.01, ***P<0.001 vs. parental sh-NC; # P<0.05, ## P<0.01 vs. spheroid sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control. SFE, spheroid formation efficiency.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Tube Formation Assay, Transwell Assay, Negative Control

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits the tumor proliferation and metastasis of HCCLM3 cells in vivo . Tumor (A) proliferation and (B) weights were measured to assess the effects of CCT2 knockdown on HCCLM3 tumor proliferation in nude mice. (C) Number of metastatic nodules stained with hematoxylin and eosin was calculated to evaluate the effects of CCT2 knockdown on the hematogenous lung metastasis of HCCLM3 cells (n=5/group). *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, In Vivo, Staining, Negative Control

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: Knockdown of CCT2 inhibits STAT3 signaling activation in hepatocellular carcinoma cells. The protein levels of STAT3, p-STAT3, MCL1, MMP2 and SOX2 in (A) Huh-7 and (B) HCCLM3 cells were measured by western blotting. The protein levels of (C) p-STAT3, (D) MCL1, (E) MMP2 and (F) SOX2 in subcutaneous tumor tissue were detected by immunohistochemical staining. *P<0.05, **P<0.01 vs. sh-NC. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; NC, negative control; p, phosphorylated; MCL1, myeloid cell leukemia sequence 1; SOX2, SRY-box transcription factor 2.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Activation Assay, Western Blot, Immunohistochemical staining, Staining, Negative Control, Sequencing

Journal: Oncology Reports

Article Title: Knockdown of CCT2 inhibits the malignant progression of hepatocellular carcinoma cells by impairing STAT3 activation

doi: 10.3892/or.2026.9086

Figure Lengend Snippet: IL-6 abolishes the effects of CCT2 knockdown on the proliferation and invasion of hepatocellular carcinoma cells. The protein levels of STAT3 and p-STAT3 in (A) Huh-7 and (B) HCCLM3 cells were measured by western blotting. The proliferation of (C) Huh-7 and (D) HCCLM3 cells following CCT2 knockdown and IL-6 treatment was detected using EdU incorporation assay. The invasion of (E) Huh-7 and (F) HCCLM3 cells was assessed using Transwell assay. *P<0.05, **P<0.01 vs. PBS. CCT2, chaperonin containing TCP1 subunit 2; sh, short hairpin; p-, phosphorylated.

Article Snippet: The primary antibodies were as follows: CCT2 (cat. no. 24896-1-AP), β-actin (cat. no. 66009-1-Ig), MMP2 (cat. no. 10373-2-AP), myeloid cell leukemia sequence 1 (MCL1; cat. no. 16225-1-AP) and SRY-box transcription factor 2 (SOX2; cat. no. 11064-1-AP; all Proteintech Group, Inc.) and STAT3 (cat. no. 4904) and phosphorylated (p-)STAT3 (Tyr705; cat. no. 4113; both Cell Signaling Technology, Inc.) The membranes were washed three times in TBST (0.1% Tween-20) for 5 min each at room temperature.

Techniques: Knockdown, Western Blot, Transwell Assay

Journal: The Journal of Biological Chemistry

Article Title: Hsp90 co-chaperone FKBP4 facilitates CCT8 folding and connects Hsp90 to chaperonin-dependent proteostasis

doi: 10.1016/j.jbc.2025.110914

Figure Lengend 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.

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 (1:500, sc-374152, Santa-cruz Biotechnology), CCT3 (1:500, sc-271336, Santa-cruz Biotechnology), HA (1:5000, MMS-101R, Covance), LexA (1:1000, sc-7544, Santa-cruz Biotechnology), and Pgk1 (1:10000, 459250, Invitrogen).

Techniques: Knockdown, TRAP Assay, Membrane, Western Blot, Expressing, Selection, Control, Transfection, Plasmid Preparation, Copurification, Comparison, Two Tailed Test