Fig 2: Knockdown of CBX3 reversed the tumor growth inhibitory effect of LINC00998.a, b Overexpression of LINC00998 inhibited A172 and U251 cell growth, while siRNA-mediated knockdown of CBX3 reversed the growth inhibitory effect of LINC00998. c Overexpression of LINC00998 inhibited A172 and U251 cell clone formation ability, while siRNA-mediated knockdown of CBX3 reversed the growth inhibitory effect of LINC00998. d Overexpression of LINC00998 inhibited A172 and U251 cell sphere formation ability, while siRNA-mediated knockdown of CBX3 reversed the growth inhibitory effect of LINC00998. e Overexpression of LINC00998 decreased the phosphorylation levels of Akt, mTOR, and P70S6K, while siRNA-mediated knockdown of CBX3 increased the phosphorylation levels of Akt, mTOR and P70S6K. (*P < 0.05, **P < 0.01, ***P < 0.001).

Fig 3: Rpap3 deletion decreases expression of R2TP clients and leads to cytoplasmic accumulation of RNA polymerase II in intestinal crypts and TA compartment.a Images are tissue sections of small intestines stained by immunohistochemistry (IHC) for Rpb1, the catalytic subunit of RNA polymerase II, from control Rpap3flox/flox mice (blue frame, left panel), or VilCreERT2; Rpap3flox/flox animals at day 6 (red frame, right panel), with magnifications of crypts (scale bars used for the two magnification insets are identical between blue and red frames). Note that the staining in stromal cells is nuclear in both wild-type and VilCreERT2; Rpap3flox/flox animals (stromal cells do not express the Cre) and control epithelium, while it becomes cytoplasmic in the mutant epithelium. Panels are representative for n = 6 animals from three independent experiments. Scale bar is 50 and 10 µm for insets, as shown in control panels. b Schematical interpretation of the micrographs in (a). In control epithelial cells (blue), R2TP incorporates Rpb1 into RNA PolII, which is then imported into the nucleus. In the absence of Rpap3 (red), neo-synthesized Rpb1 accumulates in the cytoplasm. c, d Western blot analysis of preparations enriched for epithelial crypt cells from the jejunum of animals, sacrificed 6 days after the first tamoxifen injection. NOP58, EFTUD2, and PRPF8 (c), mTOR, ATM, ATR, and TRRAP (d) were detected with specific antibodies. Tubulin and GAPDH were used as loading controls. Quantification of the signal ratios are indicated on top of each lane (average for the control ratios was arbitrarily set to 100). Each lane was loaded with the lysate obtained from one animal of the indicated genotype (n = 3 per genotype). Similar results were obtained with animals from at least two independent experiments. Apparent molecular weights are indicated on the right. Source data are provided as a “Source Data file”.

Fig 4: IL-18 stimulation induced autophagy deficiency via PI3K/Akt/mTOR signaling pathway. The chondrocytes were treated with IL-18 at different concentrations for 24 h. Protein levels of p-PI3K (B), PI3K (C), p-Akt (D), Akt (E), p-mTOR (F), mTOR (G), and GAPDH as an internal control in total extract, analyzed by Western blot (A). The values are expressed as mean ± standard deviation (SD). Significance was calculated by a one-way ANOVA with a post hoc Tukey's multiple comparisons test. *p<0.05 versus 0 ng/ml IL-18 treated group.

Fig 5: Schematic representation of a proposed mechanism of CCL5 release involving activation of the mTOR pathway. In our study, mTOR signaling is activated in ASD patients. Phosphorylated mTOR at serine residue 2448 down-regulates the phosphorylation of GSK-3ß at Ser9, and contributes to the reduction in phosphorylation of CREB at Ser133, which leads to the decreased binding of CREB to CREBBP. Thereby more available CREBBP could bind with NF-?B, which promotes the CCL5 production. The up-regulation and down-regulation of the component involved in the signaling pathway are indicated with an up arrow (red) and a down arrow (blue), respectively.