FTH1 Antibody - #DF6278

Figure 2 VDR activation suppressed ferroptosis in HG‐cultured HK‐2 cells. The mitochondrial morphology of HK‐2 cells was visualized by TEM. The arrow indicates microscopic changes in mitochondria. Absolute counting of the number of cristae per mitochondria. Scale bar = 2 or 0.5 µm (A). Quantitative analysis of iron (B), MDA (C), and GSH (D) levels in each cell group. The relative mRNA levels of SLC7A11, GPX4, TFRC,F TH1, and AIFM2 in different groups of HK‐2 cells were determined by qRT‐PCR. ACTB served as a loading control (E). Western blotting was applied to detect the protein expression levels of SLC7A11, GPX4, TFR‐1, FTH‐1, and FSP1, followed by densitometric analysis of the blots. β‐actin served as a loading control (F). Immunofluorescence staining and fluorescence intensity analysis of SLC7A11, GPX4, and TFR‐1 in different groups of HK‐2 cells as indicated. Scale bar = 20 µm (G). Each bar represents the mean ± SD of the data derived from three independent experiments (n = 3). ** p < 0.01 versus NG group; # p < 0.05, ## p < 0.01 versus HG group; & p < 0.05, && p < 0.01 versus HG group.

Fig. 5. Disordered iron homeostasis and inhibited mTORC1 activity upon exposure to CdCl2 and PCB77 at low dose. (A) The relative fluorescence intensity of CAeAM for measuring LIP to reflect intracellular iron availability (n = 3–4), and (B) Representative blots of FTH1 protein content to reflect iron storage. Analyses were performed after single or combined exposure to CdCl2 and PCB77 at 1 μM for 48 h. (C) Phosphorylated S6 and total S6 content to re- flect mTORC1 activity as measured by Western blot. Ratio of FTH1 to eIF2αP and ratio of pS6 to S6 in the control group were defined as 1. Analyses were performed after single or combined exposure to CdCl2 and PCB77 at 1 μM for 48 h. a- significantly different from the control group. Data were presented in mean ± SE. P < 0.05 was considered statistically significant.

Fig. 5 ZIP8 regulates ferroptosis of monocytes under sepsis conditions in vitro. a. Representative immunoblots of ZIP8-G, ZIP8-N, GPX4, FTH1 and xCT in RAW264.7 cells treated with different concentrations of LPS and durations. b-f. Immunoblot analyses of ZIP8-G (b, n = 4/group), ZIP8-N (c, n = 4/group), GPX4 (d, n = 4/group), FTH1 (e, n = 4/group), and xCT (f, n = 4/group) exhibited a sequential pattern of expression changes between ZIP8 (peaked at 6 h post-treatment) and ferroptosis related GPX4 and FTH1 (hit the lowest at 12 h post-treatment). g. Representative immunoblots of ZIP8-G, ZIP8-N, GPX4, FTH1 and xCT in RAW264.7 cells with/without decreased Slc39a8 expression and/or LPS treatment. h-l. Immunoblot analyses of ZIP8-G (h, n = 5/group), ZIP8-N (i, n = 5/group), GPX4 (j, n = 5/group), FTH1 (k, n = 5/group), and xCT (l, n = 5/group) exhibited that suppressing ZIP8 attenuated the decrease of GPX4, FTH1 and xCT in response to LPS. m-o. Quantification analysis showed that decreasing ZIP8 alleviated the LPS-induced upregulations of MDA (m, n = 6/group) and ferrous ions (o, n = 6/group) and reversed the decreased GSH level (n, n = 6/group). One-way ANOVA was performed in b-f. Two-way ANOVA was performed in h-o. Compared to PBS treated si-NC group:

Fig. 5| Overexpression of BCAT1 and BCAT2 ameliorated AMIinduced myocardial injury. All mice were killed 24 h after the left anterior descending CAL. The heart-specifc BCAT1 or BCAT2 over-expression mice were constructed by direct injecting Adenoassociated virus (AAV) vectors carrying BCAT1 or BCAT2 in the LV free wall in mice using a syringe with a 30-gauge needle.h Representative immunohistochemical analysis of the expression of FHC, FLC, GPX4 and Caspase-3 in heart tissue of sham and model mice with cardiac overexpression of BCAT1 and BCAT2 (n = 3 mice) (400 ×  magnifcation, scale bar, 50 µm). #p < 0.05, ##p < 0.01 versus sham group, *p < 0.05, **p < 0.01 versus model group

Fig. 3. YAP1 knockout accelerates ferroptosis in PM2.5-induced lung toxicity. A, Representative fluorescent images of ROS staining; B, Quantification of ROS labeling intensity in the lung; C, Representative TEM images. The red arrow indicates representative mitochondria in WT or YAP1-CKO mouse lungs exposed to PM2.5 or not; D, Immunoblotting detection of SLC7A11, GPX4 and FTH1 expression in mice with the indicated treatment; quantitative results are shown in E-G n = 5; lung content of Fe2+ (H), MDA (I) and GSH (J) measured by the corresponding commercial kit, n = 5. The values are shown as the mean ± SD,