rfc3  (Proteintech)

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 1. RFC3 displays upregulated expression in CRC cells. (A) RFC3 expression in normal and CRC tumor tissues of patients was analyzed using the TNMplot tool. (B) Box plot illustrates RFC3 expression level in 275 COAD tissues (red) vs. normal 349 tissues (gray) derived from the GEPIA database. *P<0.05. (C) Reverse transcription‑quantitative PCR and (D) western blotting analysis of RFC3 expression. ***P<0.001 vs. HIEC‑6 group. CRC, colorectal cancer; RFC3, replication factor C subunit 3; COAD, colon adenocarcinoma.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Expressing, Derivative Assay, Western Blot

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 2. RFC3 depletion diminishes the proliferation of SW620 cells. (A) Reverse transcription‑quantitative PCR and (B) western blotting examined the transfection efficacy of siRNA‑RFC3‑1/2. (C) Cell viability was assessed using the Cell Counting Kit‑8 method. (D) EDU staining (magnification, x200) measured cell proliferation. **P<0.01, ***P<0.001 vs. siRNA‑NC group. RFC3, replication factor C subunit 3; siRNA, small interfering RNA; NC, negative control.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Western Blot, Transfection, CCK-8 Assay, Staining, Small Interfering RNA, Negative Control

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 3. RFC3 depletion aggravates the apoptosis of SW620 cells. (A) Flow cytometric analysis of cell apoptosis and (B) quantification. (C) Western blot‑ ting tested the expression of apoptosis‑associated proteins. **P<0.01, ***P<0.001 vs. siRNA‑NC group. RFC3, replication factor C subunit 3; siRNA, small interfering RNA; NC, negative control.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Western Blot, Expressing, Small Interfering RNA, Negative Control

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 4. RFC3 depletion obstructs SW620 cell migration, invasion and angiogenesis. (A) Wound healing (magnification, x100) and (B) Transwell assays (magnification, x200) estimated the migration and invasion of cells. (C) Western blotting tested the expression of metastasis‑associated proteins. ***P<0.001 vs. siRNA‑NC group. (D) Tube formation assay (magnification, x100) assessed HUVECs angiogenesis in the conditioned medium of SW620 cells. (E) Western blotting tested the expression of angiogenesis‑associated proteins. **P<0.01 vs. siRNA‑NC (CM) + HUVEC group. RFC3, replication factor C subunit 3; siRNA, small interfering RNA; NC, negative control; MMP, matrix metallopeptidase; VEGF, vascular epidermal growth factor; VEGFR, VEGF receptor.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Migration, Western Blot, Expressing, Tube Formation Assay, Small Interfering RNA, Negative Control

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 5. RFC3 protein interacts with KIF14 protein. (A) Coexpedia database predicted the co‑expression of RFC3 and KIF14. (B) Biogrid database predicted the relationship between RFC3 and KIF14. (C) KIF14 expression in normal and CRC tumor tissues of patients was analyzed using the TNMplot tool. (D) The box plot illustrated KIF14 expression level in 275 COAD tissues vs. normal 349 tissues derived from the GEPIA database. *P<0.05. (E) Long‑term survival between patients with low and high KIF14 expression levels, according to KM‑plot database. (F) Western blotting of KIF14 expression after RFC3 was depleted. ***P<0.001 vs. siRNA‑NC group. (G) Co‑immunoprecipitation assay validated the interaction between RFC3 and KIF14. RFC3, replication factor C subunit 3; siRNA, small interfering RNA; NC, negative control; KIF14, kinesin family member 14; CRC, colorectal cancer; COAD, colon adenocarcinoma; GEPIA, Gene Expression Profiling Interactive Analysis.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Expressing, Derivative Assay, Western Blot, Co-Immunoprecipitation Assay, Small Interfering RNA, Negative Control, Gene Expression

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 6. Downregulation of KIF14 mediated by RFC3 insufficiency decreases the proliferation of SW620 cells. (A) Reverse transcription‑quantitative PCR and (B) western blotting examined the transfection efficacy of Ov‑KIF14 plasmids. ***P<0.001 vs. Ov‑NC group. (C) Cell viability was assessed using the Cell Counting Kit‑8 method. (D) EDU staining (magnification, x200) measured cell proliferation. ***P<0.001 vs. siRNA‑NC group; ###P<0.001 vs. siRNA‑RFC3 + Ov‑NC group. RFC3, replication factor C subunit 3; Ov, overexpression; NC, negative control; KIF14, kinesin family member 14.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Western Blot, Transfection, CCK-8 Assay, Staining, Over Expression, Negative Control

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 7. Downregulation of KIF14 mediated by RFC3 insufficiency increases the apoptosis of SW620 cells. (A) Flow cytometric analysis of apoptosis and (B) quantification. (C) Western blotting tested the expression of apoptosis‑associated proteins. ***P<0.001 vs. siRNA‑NC group; #P<0.05, ##P<0.01, ###P<0.001 vs. siRNA‑RFC3 + Ov‑NC group. RFC3, replication factor C subunit 3; Ov, overexpression; NC, negative control; KIF14, kinesin family member 14.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Western Blot, Expressing, Over Expression, Negative Control

Journal: Experimental and therapeutic medicine

Article Title: RFC3 drives the proliferation, migration, invasion and angiogenesis of colorectal cancer cells by binding KIF14.

doi: 10.3892/etm.2024.12510

Figure Lengend Snippet: Figure 8. Downregulation of KIF14 mediated by RFC3 insufficiency decreases the migration, invasion and angiogenesis of SW620 cells. (A) Wound healing (magnification, x100) and (B) Transwell assays (magnification, x200) estimated the migration and invasion of cells. (C) Western blotting tested the expression of metastasis‑associated proteins. ***P<0.001 vs. siRNA‑NC group; ##P<0.01, ###P<0.001 vs. siRNA‑RFC3 + Ov‑NC group. (D) Tube formation assay (magnifi‑ cation, x100) assessed HUVEC angiogenesis in the conditioned medium of SW620 cells. (E) Western blotting tested the expression of angiogenesis‑associated proteins. ***P<0.001 vs. siRNA‑NC (CM) + HUVEC group; ##P<0.01 vs. siRNA‑RFC3 + Ov‑NC (CM) + HUVEC group. RFC3, replication factor C subunit 3; Ov, overexpression; NC, negative control; KIF14, kinesin family member 14; siRNA, small interfering RNA; VEGF, vascular epidermal growth factor; VEGFR, VEGF receptor.

Article Snippet: Next, 2 μg RFC3 (cat. no. 11814‐1‐AP; Proteintech Group, Inc.), KIF14 (cat. no. ab71155; Abcam) or goat anti‐rabbit IgG (the negative control; cat. no. ab172730; Abcam) antibodies were added to 500 μg SW620 cell lysates and incubated over‐ night at 4 ̊C.

Techniques: Migration, Western Blot, Expressing, Tube Formation Assay, Over Expression, Negative Control, Small Interfering RNA