Display Settings:

Items per page

Results: 8

1.
Figure 6

Figure 6. No significant changes were observed in the membrane bound TNF-α protein after UVB and IL-1α treatment. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Cells were treated with sham, sham + IL-1α, UVB, and UVB+IL-1α for various time points. Membrane fractions were eluted and an equal amount of protein was used for TNF-α ELISA.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.
2.

Figure 5. Synergistic increase of TNF-α protein in keratinocyte medium corresponds to TNF-α mRNA increase due to UVB and IL-1α response. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Cells were treated with sham, sham + IL-1α, UVB, and UVB+IL-1α for various time points. Supernatant samples were collected from sham, sham + IL-1α, UVB, and UVB+IL-1α-treated cells at the indicated time points. An equal amount of protein was used for TNF-α ELISA.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.
3.
Figure 4

Figure 4. Exogenous rTNF-α increases TNF-α mRNA induction in UVB-irradiated neonatal keratinocytes. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Cells were treated with sham, sham + rTNF-α (5ng/ml), UVB, and UVB + rTNF-α. RNA sample were collected 3h post irradiation and analyzed by real-time PCR. Results were normalized to PPIA (housekeeping gene control). Expression levels of TNF-α mRNA are indicated as “fold change” compared to control.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.
4.
Figure 3

Figure 3. IFN-α2b has no effect on TNF-α mRNA synthesis in sham- or UVB-irradiated keratinocytes in the presence or absence of IL-1α. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Sham- or UVB-irradiated keratinocytes were treated with IFN-α2b (2000u/ml) or IL-1α (10 ng/ml) alone or in combination with IFN-α2b + IL-1α. RNA sample were collected after 6 h and analyzed by real-time PCR. Results were normalized to PPIA mRNA and compared to control groups.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.
5.
Figure 8

Figure 8. TNF-α mRNA stability is not induced after UVB irradiation and IL-1α stimulation. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Keratinocytes were UVB or sham-irradiated and stimulated with IL-1α for 3 h. Actinomycin D (10 μg/ml) was added after 3h. Total RNA was extracted at different time points and TNF-α mRNA levels were determined by northern analysis and Real-time PCR. The half-life was calculated in UVB, and UVB+ IL-1α treated cells.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.
6.
Figure 7

Figure 7. UVB irradiation and IL-1α enhances TNF-α gene transcription. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Nuclei were isolated 3 h after sham- or UVB-irradiation and stimulated with or without IL-1α. Nuclear run-off assay was performed to examine TNF-α and 18S (control) transcription. Detectable bands were assessed by densitometric analysis. UVB irradiation induced TNF-α gene transcription 5-fold. One of four experiments is shown, illustrating 11-fold induction of UVB+ IL-1α cells when compared with sham irradiated cells.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.
7.

Figure 1. IL-1α synergistically increases TNF-α mRNA induction by UVB-irradiated neonatal KCs. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Cells were treated with sham, sham+IL-1α (10 ng/ml), UVB (30mJ/cm2) and UVB+IL-1α. RNA samples were collected at the indicated times and analyzed by real-time PCR. TNF-α mRNA cycle numbers were normalized to cyclophilin A (PPIA). Expression levels of TNF-α mRNA are indicated as “fold change” compared to control. UVB+IL-1α-treated cells showed synergistically increased TNF-α mRNA as compared to UVB or IL-1α alone (Figure 1a and b). The time-course study showed that there was no biphasic induction of TNF-α mRNA in keratinocytes up to 72h.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.
8.
Figure 2

Figure 2. Anti-TNF-α antibody has no effect on TNF-α mRNA synthesis in UVB-irradiated keratinocytes in the presence of IL-1α. From: UVB and pro-inflammatory cytokines synergistically activate TNF-? production in keratinocytes through enhanced gene transcription.

Controls included cells treated with sham, sham+IL-1α, UVB, and UVB+IL-1α. In the test group, UVB+ IL-1α-treated cells were also incubated with anti-TNF-α antibody (5, 10, 100 ng/ml). RNA samples were collected after 3h and analyzed by real-time PCR. Results were normalized to PPIA mRNA and compared to control groups. No significant difference in expression of TNF-α mRNA is observed between UVB+ IL-1α and UVB+ IL-1α + anti-TNF-α treated keratinocytes.

Muhammad M. Bashir, et al. J Invest Dermatol. ;129(4):994-1001.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk