primary antibody rabbit igg polyclonal abca1  (Thermo Fisher)

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Macrophage migration inhibitory factor is regulated by HIF-1α and cAMP and promotes renal cyst cell proliferation in a macrophage-independent manner

doi: 10.1007/s00109-020-01964-1

Figure Lengend Snippet: MIF and ABCA1 are expressed in a HIF-1α-dependent manner in cyst-lining cells of an ADPKD mouse model. Tamoxifen was applied at postnatal days 35–37 to induce tubule-specific deletion of PKD1 in Ksp CreER T2 ; Pkd1 lox;lox ( Pkd1 fl;fl ; n = 7) mice. In parallel, genetic deletion was induced in Ksp CreER T2 ; Pkd1 lox;lox ;Hif-1α lox/lox ( Pkd1 fl;fl ; Hif-1α fl;fl ; n = 5) mice to receive tubular codeletion of PKD1 and HIF-1α. Mice were then either treated with the prolylhydroxylase inhibitor 2-(1-chloro-4-hydroxyisoquinoline-3-carboxamido) acetate ( Pkd1 fl;fl + ICA; n = 6); ( Pkd1 fl;fl ; Hif-1α fl;fl + ICA; n = 6) or its vehicle for 12 weeks. Noninduced mice served as controls (Ctrl; n = 4). a As shown previously, the abovementioned ADPKD mouse model ( Pkd1 fl;fl ) shows a mild progression which does not lead to hypoxia or induction of HIF-1α. In line with these findings, MIF expression did not differ in the cortex between Ctrl, Pkd1 fl;fl , and Pkd1 fl;fl ; Hif-1α fl;fl kidneys. However, application of ICA ( Pkd1 fl;fl + ICA) resulted in a significant increase of HIF-1α shown previously which was prevented in mice co-deleted for HIF-1α ( Pkd1 fl;fl ; Hif-1α fl;fl + ICA). In line with the assumption of MIF being regulated by HIF-1α, MIF expression was significantly increased in the cortex of Pkd1 fl;fl + ICA mice which could be prevented in mice codeleted for HIF-1α ( Pkd1 fl;fl ; Hif-1α fl;fl + ICA). b ABCA1 shows a comparable pattern of HIF-dependent expression as MIF in cyst cells in the renal cortex of the chosen models. *Significant compared with Ctrl. §Significant compared with Pkd1 fl;fl + ICA

Article Snippet: Sections were incubated in protein block serum free (X0909, Dako) for 60 min. Primary antibody rabbit IgG polyclonal ABCA1 (1:500, Thermo Fisher Scientific, Inc.) was incubated at 4 °C overnight.

Techniques: Expressing

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Macrophage migration inhibitory factor is regulated by HIF-1α and cAMP and promotes renal cyst cell proliferation in a macrophage-independent manner

doi: 10.1007/s00109-020-01964-1

Figure Lengend Snippet: MIF and ABCA1 are HIF-1α target genes in human primary tubular epithelial cells (hPTECs). a HIF stabilization by DMOG in hPTCs for 16 h led to a significantly increased expression of MIF and ABCA1 mRNA ( n = 13 for MIF, n = 8 for ABCA1, and n = 6 for EGLN3, egl nine homolog 3) compared with vehicle control. The known HIF-1α target gene EGLN3 served as positive control. b HIF-1α- and HIF-1β ChIP-seq signals at the genomic regions coding for MIF or ABCA1, respectively, reveal HIF DNA interactions at the promoter of MIF and at two regions at the ABCA1 locus (dotted rectangles) in hPTCs. MIF-AS1: MIF antisense 1 which is encoded close to MIF in the indicated genomic interval of the human genome. c Expression qPCR of MIF and ABCA1, respectively, in HIF-1α and HIF-1β siRNA depleted hPTCs with or without DMOG treatment for 16 h. MIF or ABCA1 mRNA levels increased in DMOG-treated cells and are significantly reduced in HIF-depleted cells. EGLN3 served as positive control ( n = 3 individual experiments). *Significant compared with vehicle control. §Significant compared with cells treated with nontargeting (nt) siRNA

Article Snippet: Sections were incubated in protein block serum free (X0909, Dako) for 60 min. Primary antibody rabbit IgG polyclonal ABCA1 (1:500, Thermo Fisher Scientific, Inc.) was incubated at 4 °C overnight.

Techniques: Expressing, Control, Positive Control, ChIP-sequencing