antibody against ca9  (Bio-Techne corporation)

Journal: Cancer Medicine

Article Title: IGFL2‐AS1 ‐induced suppression of HIF ‐1α degradation promotes cell proliferation and invasion in colorectal cancer by upregulating CA9

doi: 10.1002/cam4.5562

Figure Lengend Snippet: IGFL2‐AS1 positively regulates CA9 expression in CRC. (A, B) Differential gene heat map (A) and volcano map (B) of negative control and IGFL2‐AS1 knockdown LoVo cell populations. Upregulation is shown as red, and downregulation is shown as green. (C) mRNA levels of 9 downregulated genes in IGFL2‐AS1 knockdown LoVo cells were measured by qRT‐PCR. (D, E) qRT‐PCR and western blotting were utilized to detect mRNA (D) and protein (E) levels of CA9 in HT29 and LoVo cells that overexpress or knockdown IGFL2‐AS1. (F) The expression level of CA9 in normal colonic epithelial cell line and CRC cell lines were detected by qPCR and western blotting. (G) Immunohistochemistry was performed to assess the protein level of CA9 in CRC tumor tissue and adjacent non‐tumor tissue. (H) CA9 expression in TCGA‐derived specimen datasets in the GEPIA database. (magnification, 200× and 400×; scale bar, 100 and 50 μm; *p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The slices were incubated with specific primary antibodies against CA9 (1:2000, ab243660, Abcam, Cambridge, UK), HIF‐1α (1:300, 66,730‐1‐Ig, Proteintech, Wuhan, China), and Ki67 (1:500, ab92742, Abcam, Cambridge, UK) overnight (14–16 h) at 4°C.

Techniques: Expressing, Negative Control, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Derivative Assay

Journal: Cancer Medicine

Article Title: IGFL2‐AS1 ‐induced suppression of HIF ‐1α degradation promotes cell proliferation and invasion in colorectal cancer by upregulating CA9

doi: 10.1002/cam4.5562

Figure Lengend Snippet: CA9 is required for IGFL2‐AS1 to enhance CRC cell growth, migration, and invasion in vitro. (A) The effect of CA9 regulated by IGFL2‐AS1 on CRC cell viability was detected by CCK‐8 assay. (B, C) The quantitative results and images of the EdU assay showing that IGFL2‐AS1 regulates CRC cell proliferation through CA9. (D) The effect of CA9 overexpression and IGFL2‐AS1 knockdown co‐transfection on CRC cell proliferation was examined using a colony formation assay. (E, F) The effect of migration and invasion of IGFL2‐AS1 and CA9 co‐transfection in LoVo cells was determined by transwell assay. (magnification, 200× and 400×; scale bar, 25 μm; * p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The slices were incubated with specific primary antibodies against CA9 (1:2000, ab243660, Abcam, Cambridge, UK), HIF‐1α (1:300, 66,730‐1‐Ig, Proteintech, Wuhan, China), and Ki67 (1:500, ab92742, Abcam, Cambridge, UK) overnight (14–16 h) at 4°C.

Techniques: Migration, In Vitro, CCK-8 Assay, EdU Assay, Over Expression, Cotransfection, Colony Assay, Transwell Assay

Journal: Cancer Medicine

Article Title: IGFL2‐AS1 ‐induced suppression of HIF ‐1α degradation promotes cell proliferation and invasion in colorectal cancer by upregulating CA9

doi: 10.1002/cam4.5562

Figure Lengend Snippet: IGFL2‐AS1 upregulates CA9 expression by inhibiting HIF‐1α proteolysis. (A) Western blotting was used to assess the HIF‐1α and CA9 expression levels in CRC cells with IGFL2‐AS1 knockdown and overexpression. (B) The effect of HIF‐1α knockdown on the increase in CA9 protein level by IGFL2‐AS1 overexpression. (C) The protein levels of HIF‐1α after HT29 and LoVo cells were treated with MG132 (10 μM) for 0, 3, 6, and 9 h and were measured by western blotting. (D) CRC cells were transfected with sh IGFL2‐AS1#1, oe IGFL2‐AS1, or negative control, and treated with or without MG132 (10 μM) for 6 h. The protein levels of HIF‐1α were detected by western blotting. (E) CRC cells in IGFL2‐AS1 knockdown and overexpression groups were treated with CHX (100 μg/mL) for 0, 3, 6, and 9 h, and western blotting was performed to measure the HIF‐1α expression levels

Article Snippet: The slices were incubated with specific primary antibodies against CA9 (1:2000, ab243660, Abcam, Cambridge, UK), HIF‐1α (1:300, 66,730‐1‐Ig, Proteintech, Wuhan, China), and Ki67 (1:500, ab92742, Abcam, Cambridge, UK) overnight (14–16 h) at 4°C.

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

Journal: Cancer Medicine

Article Title: IGFL2‐AS1 ‐induced suppression of HIF ‐1α degradation promotes cell proliferation and invasion in colorectal cancer by upregulating CA9

doi: 10.1002/cam4.5562

Figure Lengend Snippet: IGFL2‐AS1 accelerates CRC tumor growth in vivo. (A, B) All mice were sacrificed on day 27 after injection, and tumors were harvested and photographed. (C) The histogram of tumor weight and the tumor growth curve is based on measured tumor volume. (D) Western blotting was performed to verify the protein levels of HIF‐1α and CA9 in the tumors of nude mice in each group. Two samples were taken from each group. (E) Immunohistochemical staining of HIF‐1α, CA9, and Ki67 in xenograft tumors. (scale bar, 1 cm; * p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The slices were incubated with specific primary antibodies against CA9 (1:2000, ab243660, Abcam, Cambridge, UK), HIF‐1α (1:300, 66,730‐1‐Ig, Proteintech, Wuhan, China), and Ki67 (1:500, ab92742, Abcam, Cambridge, UK) overnight (14–16 h) at 4°C.

Techniques: In Vivo, Injection, Western Blot, Immunohistochemical staining, Staining

Journal: Cancer Medicine

Article Title: IGFL2‐AS1 ‐induced suppression of HIF ‐1α degradation promotes cell proliferation and invasion in colorectal cancer by upregulating CA9

doi: 10.1002/cam4.5562

Figure Lengend Snippet: The schematic illustration of a model for IGFL2‐AS1 to promote tumor growth in CRC. IGFL2‐AS1 is upregulated in CRC and may inhibit the proteasomal degradation of HIF‐1α, which could increase HIF‐1α expression and its downstream gene CA9, thereby promoting CRC cell proliferation, migration, and invasion

Article Snippet: The slices were incubated with specific primary antibodies against CA9 (1:2000, ab243660, Abcam, Cambridge, UK), HIF‐1α (1:300, 66,730‐1‐Ig, Proteintech, Wuhan, China), and Ki67 (1:500, ab92742, Abcam, Cambridge, UK) overnight (14–16 h) at 4°C.

Techniques: Expressing, Migration