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1.
Figure 2

Figure 2. Fold change in RNA levels of cartilage from ischemic and control group.. From: Regulation of VEGF expression by HIF-1? in the femoral head cartilage following ischemia osteonecrosis.

RNA was isolated from cartilage 24 hr after ischemic induction in pig model. RNA levels were measured by real-time RT-PCR. Level of RNA from control group was normalized to a value of 1.

Chi Zhang, et al. Sci Rep. 2012;2:650.
2.
Figure 1

Figure 1. The vessel formation increased after ischemia induction in pig femoral head.. From: Regulation of VEGF expression by HIF-1? in the femoral head cartilage following ischemia osteonecrosis.

The femoral heads were obtained two weeks following the surgery to induce femoral head ischemia. Histological section of the epiphyseal cartilage from the control (a) and the ischemic sides (b) demonstrating increased vessel formation in cartilage in the ischemic side.

Chi Zhang, et al. Sci Rep. 2012;2:650.
3.
Figure 6

Figure 6. Upregulation of VEGF during hypoxia is mediated partially through HIF-1α.. From: Regulation of VEGF expression by HIF-1? in the femoral head cartilage following ischemia osteonecrosis.

RNA expression levels of HIF-1α (a) and VEGF (b) as determined by quantitative real-time RT-PCR. HEK293 cells were transfected with siRNA targeting HIF-1α, and then cultured in hypoxia station. RNA was isolated in 24 hr and quantitated by real-time RT-PCR. The RNA level from the control group was normalized to a value of 1. Values were presented as the mean ±S.D.

Chi Zhang, et al. Sci Rep. 2012;2:650.
4.
Figure 3

Figure 3. HIF-1α expression increased in chondrocytes of ischemic group.. From: Regulation of VEGF expression by HIF-1? in the femoral head cartilage following ischemia osteonecrosis.

HIF-1α immunostaining of epiphyseal cartilage two weeks after the surgery to induce ischemia. Increased immunostaining was observed in the ischemic side compared to the control side. Higher magnifications of control (a) and ischemic (b) are shown. Cells were counted at 40x magnification. At least 300 cells were counted and expressed as the percent of positive cells (c). A paired t-test was performed comparing the control and the contralateral ischemic sides.

Chi Zhang, et al. Sci Rep. 2012;2:650.
5.
Figure 4

Figure 4. VEGF expression increased in chondrocytes of ischemic group.. From: Regulation of VEGF expression by HIF-1? in the femoral head cartilage following ischemia osteonecrosis.

VEGF immunostaining of epiphyseal cartilage two weeks after the surgery to induce ischemia. Increased immunostaining was observed in the ischemic side compared to the control side. Higher magnifications of control (a) and ischemic (b) are shown, respectively. Cells were counted at 40x magnification. At least 300 cells were counted and expressed as the percent of positive cells (c). A paired t-test was performed comparing the control and the contralateral ischemic sides.

Chi Zhang, et al. Sci Rep. 2012;2:650.
6.
Figure 5

Figure 5. Fold change in RNA levels of chondrocytes under hypoxia.. From: Regulation of VEGF expression by HIF-1? in the femoral head cartilage following ischemia osteonecrosis.

Primary chondrocytes were isolated from cartilage of pig femoral heads. Pig primary chondrocytes were cultured for 24 hr in hypoxia station with 1% or 20% of O2. RNA levels were isolated and measured by real-time RT-PCR for expression of VEGF and HIF-1α (a). Level of each type of RNA from control group was normalized to a value of 1. (b) DFO enhanced VEGF expression under hypoxia. Pig primary chondrocytes were cultured in normal (21%O2) and hypoxia (1%O2) condition. In one group of hypoxia condition, HIF-1α activator DFO was added at 100 µM. RNA was isolated 24 hr after hypoxia. RNA levels were measured by real-time RT-PCR. Level of each type of RNA from control group was normalized to a value of 1. *: A star indicates statistical significance compared to expression level in normal condition (p<0.05, N = 3). **: Two stars indicate statistical significance compared to expression level in hypoxia condition in the absence of DFO (p<0.05, N = 3).

Chi Zhang, et al. Sci Rep. 2012;2:650.

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