Rabbit anti human β actin

The following antibodies were employed in this study: rabbit anti‐human TRPC6 (Alomone Labs, Jerusalem, Israel); rabbit anti‐human podocin (Abcam, Cambridge, UK) for immunocytochemistry and another one from Santa Cruz (Heidelberg, Germany) for Western blotting; mouse anti‐human synaptopodin (Progen, Heidelberg, Germany); rabbit anti‐human PKCα, rabbit anti‐human PKCδ and rabbit anti‐human PKCζ (Biomol Res Lab, Plymouth Meeting, PA, USA); rabbit anti‐human PKCλ/ι (Santa Cruz); rabbit anti‐human PKCβ₁, rabbit anti‐human PKCβ₂, rabbit anti‐human PKCγ, rabbit anti‐human PKCε, rabbit anti‐human PKCη, rabbit anti‐human PKCθ and rabbit anti‐human β‐actin (Sigma‐Aldrich).

RNA was isolated after lysis by PeqGold TriFast (Peqlab) followed by chloroform-phenol extraction. Reverse transcription was performed with M-MLV reverse transcriptase (Thermo Fisher) and random hexamer primers (Carl Roth) as described previously (8 (link)). Quantitative real-time PCR was performed with SoFast Eva Green master mix (BioRad) and primers, the sequences of which are listed in Supplemental Table 6. Hprt (hypoxanthine guanine phosphoribosyl transferase, Entrez Gene ID 15452) served as the house-keeping gene for expression normalization. Relative gene expression was calculated with the ΔCt method and presented as log₂ expression, where log₂ expression_gene = Ct_gene — Ct_Hprt.
Protein extraction and Western blotting were performed exactly as described previously (61 (link)). We used the following antibodies: rabbit anti-mouse FTH1 (1:1000; Cell Signaling, 3998), rabbit anti-mouse Ferritin light chain (1:1000; Abcam, ab69090), mouse anti-human Transferrin receptor (1:1000; Invitrogen, PA1-84854), rabbit anti-human FPN1 (1:2000; Eurogentec, NRU 451443), and rabbit anti-human β-actin (1:500; Sigma-Aldrich, A2066).