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. 2019 Oct 11;13:446. doi: 10.3389/fncel.2019.00446

FIGURE 5.

FIGURE 5

JAK1 and ATF4 cooperatively regulate a subset of ER stress-induced genes. (A) List of genes that are both JAK1 and ATF4 dependent in response to ER stress as determined by RNA-seq and reported by others. (B) Astrocytes were transfected with control (CTL) or ATF4-targeting siRNA for 48 h and treated with thapsigargin for 4 h. Indicated gene expression was analyzed by RT-qPCR. UT = untreated. (C) Astrocytes were transfected with CLT or one of two JAK1 siRNAs and immunoblotted for ATF4. Immunoblots were quantified and normalized to GAPDH expression. (D) Astrocytes were transfected with CTL or JAK1 siRNA for 48 h and then treated with thapsigargin for 4 h. Cytosolic and nuclear fractions were isolated and analyzed by immunoblot. (E) Primary astrocytes were treated with thaps (1 μM) for 4 h. Protein lysates reserved or immunoprecipiated with α-ATF4 antibody before immunoblotting. (F) Quantification of JAK1 co-immunoprecipiated with ATF4 as shown in the top panel of (E). Data are represented as means ± standard deviation. N = 3. ∗p ≤ 0.05.